Cephalosporin antibiotics

ABSTRACT

The present invention includes novel compounds of formula ##STR1## where G, H, J, L and M are carbon or nitrogen, R 99  is selected from the group consisting of sulfur, SO, SO 2 , NH, N-alkyl, oxygen, C.tbd.C (cis or trans), and C.tbd.C, and R 12  is NR 13  R 14 , ##STR2## The invention also includes the pharmacologically acceptable salts which exhibit antibiotic activity against a wide spectrum of organisms including organisms which are resistant to β-lactam antibiotics and are useful as antibacterial agents. The invention also relates to novel intermediates useful for making the novel compounds of the present invention and to novel methods for producing the novel compounds and intermediate compounds.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel cephalosporin antibiotics andtheir methods of production and use. These compounds are expected toexhibit antibiotic activity against a wide spectrum of organisms,including organisms which are resistant to conventional β-lactamantibiotics.

2. Review of the Background Art

The following review of the background of the invention is merelyprovided to aid in the understanding of the present invention andneither it nor any of the references cited within it are admitted to beprior art to the present invention.

Over the past three decades a large variety of antibiotics have becomeavailable for clinical use. One class of antibiotics which has seenremarkable growth are the cephalosporins, over 70 of which have enteredclinical use for the treatment of bacterial infections in mammals since1965. The cephalosporins exhibit their antibacterial activity byinhibiting bacterial peptidoglycan biosynthesis, and have been extremelyeffective in treating a wide variety of bacterial infections.Cephalosporins that are said to have antibacterial activity aredescribed in U.S. Pat. Nos. 3,992,377 and 4,256,739.

Unfortunately, the wide-spread and indiscriminant use of theseantibiotics has led to a rapid increase in the number of bacterialstrains which are resistant to these compounds. Most importantly, thisresistance has emerged among clinically important microorganisms whichthreaten to limit the utility of presently available cephalosporinantibiotics. In particular, resistant strains of Salmonella, S.pneumoni.ae butted., Enterobacteriac.ae butted., and Pseudomonas haveemerged which threaten to undo many of the strides made in reducingmortality and morbidity from bacterial infections.

Bacterial resistance to cephalosporins follows three major pathways: (a)the development of β-lactamases capable of inactivating the β-lactamring of the cephalosporin; (b) decreased cephalosporin penetration intothe bacteria due to changes in bacterial cell wall composition; and (c)poor binding to penicillin-binding proteins (PBPs). The latter pathwayis especially important, as the binding of β-lactams to PBPs isessential for inhibiting bacterial cell-wall biosynthesis. CertainGram-positive bacteria, namely methicillin-resistant Staphylococcusaureus ("MRSA") and Enterococci are highly resistant to β-lactamantibiotics. Resistance in MRSA is due to the presence of high levels ofan unusual PBP, PBP2a, which is insensitive, or binds poorly, toβ-lactam antibiotics. The activity of β-lactam antibiotics againstPBP2a-containing organisms has been shown to correlate well with thebinding affinity of the antibiotic to PBP2a. Currently, theglycopeptides vancomycin and teicoplanin are primarily used for MRSAbacteremia. The quinolone antibacterials and some carbapenems, such asimipenem, have been reported to be active against a few MRSA strains,but their use is restricted due to emerging resistant MRSA strains.

Experimental compounds which may possess utility as anti-MRSA oranti-enterococcal bactericides include the glycylcyclines (see, e.g.,P.-E. Sum et al., J. Med. Chem., 37, (1994)), FK-037 (see, e.g., H. Ohkiet al., J. Antibiotics, 46: 359-361 (1993)), RP-59,500 (see, e.g., S. K.Spangler et al., Antimicro. Agents Chemother., 36: 856-9 (1992)), theeverninomycin complex (see, e.g., W. E. Sanders et al., Antimicro.Agents Chemother., 6: 232-8 (1974)), the 2-(biaryl)carbapenems (see,e.g., U.S. Pat. No. 5,025,006), 3-(benzothiazolylthio)cephems (see,e.g., EP Application No. 527686), 3-(thiazolylthio)carbacephems (see,e.g., R. J. Ternansky et al., J. Med. Chem., 36: 1971 (1993) and U.S.Pat. No. 5,077,287) and arbekacin (S. Kondo, et al. J. Antibiotics 46:531 (1993).

Recent advances in the compounds, compositions and methods useful fortreating infections in mammals arising from β-lactam antibioticresistant bacteria are described in commonly owned InternationalPublication WO 95/26966 and U.S. patent applications Ser. Nos.08/222,262, filed Apr. 1, 1994, now abandoned; 08/369,798, filed Jan. 6,1995, now abandoned; 08/413,713, now U.S. Pat. No. 5,756,493, issued May26, 1998; 08/413,714, now U.S. Pat. No. 5,607,926, issued Mar. 4, 1997;08/415,065, filed Mar. 29, 1995, now abandoned; 08/413,712, now U.S.Pat. No. 5,604,218, issued Feb. 18, 1997; 08/415,064, now U.S. Pat. No.5,789,584, issued Aug. 4, 1998; 08/415,069, now U.S. Pat. No. 593,986issued Jan. 14, 1997; 08/455,969, now U.S. Pat. No. 5,698,547, issuedDec. 16, 1997; and 08/457,673, now U.S. Pat. No. 5,688,786, issued Nov.18, 1997; all of which are incorporated herein by reference in theirentirety, including any drawings.

SUMMARY OF THE INVENTION

The present invention includes compounds, compositions and methodseffective to treat infections in mammals arising from β-lactamantibiotic resistant bacteria. Preferred compounds will have a minimuminhibitory concentration (MIC) that is less that 50%, more preferablyless than 10%, and most preferably less than 1% of the MIC of cefotaximeor imipenem for a beta-lactam resistant organism, preferably amethicillin-resistant Staphylococcal or ampicillin-resistantEnterococcal organism. Other preferred compounds will be able to preventor reduce mortality in mice infected with the beta-lactam resistantorganism to a greater extent than cefotaxime or imipenem.

In one aspect the invention features compounds of the structures II--VI##STR3## and/or their pharmaceutically acceptable salts and/or prodrugs,wherein R¹ is selected from the group consisting of --NHC (O) ZR³, --NR⁴R⁵, and ##STR4## Z is selected from the group consisting of --CH₂(X)_(m) --, --C (NOR⁶)--, --CH(OR⁷)--, --C (CHCO₂ R⁸)-- and --CH (NR⁹R¹⁰)--;

X is selected from the group consisting of oxygen and sulphur;

m is selected from the group consisting of 0 and 1;

R³ is selected from the group consisting of cyano, alkyl, aryl,heterocycle (wherein said heterocycle is optionally substituted andpreferably is disubstituted with NH₂ and halogen (preferably chlorine)),heteroaralkyl and (CH₂)_(n) T,.

n is 1 to 6,

T is selected from the group consisting of amino, amidino (C- orN-linked), guanidino, and isothioureido, optionally substituted byalkyl, aryl, hydroxyl, or amino;

R⁴⁻⁷ are independently selected from the group consisting of hydrogen,alkyl, aryl and acyl;

R⁸ is selected from the group of hydrogen, alkyl and aryl;

R⁹ and R¹⁰ are selected independently from the group consisting ofhydrogen, alkyl, acyl, and heterocyclecarbonyl;

R² is selected from the group consisting of hydrogen, alkyl, alkenyl,aryl, heterocycle, aralkyl, heteroaralkyl, and trialkylsilyl; or R² isnot present and the CO₂ group to which it would be attached bears anegative charge.

A, B, D, and E are selected from the group consisting of carbon,nitrogen and sulphur and the specific juxtaposition of groups A, B, Dand E is limited to examples of heterocyclic groups known in thechemistry arts;

G, H, J, L and M are carbon, nitrogen or ⁺ NR¹¹ (quaternary ammoniumheterocycle) and the specific juxtaposition of groups G, H, J, L and Mis limited to examples of heterocyclic groups known in the chemistryarts;

R¹¹ is selected from the group consisting of H, halogen, alkyl, alkoxy,hydroxyl, amino, cyano, hydroxyalkyl, carboxamidoalkyl, optionallysubstituted aminoalkyl or quaternary ammonium alkyl, and quaternaryheteroaryliumalkyl;

alk₁ and alk₂ are alkyl groups and are independently and optionallysubstituted with a substitutent selected from the group consisting ofalkyl, hydroxyl, optionally substituted amino, alkoxy, hydroxyalkyl andoptionally substituted carboxamide;

p is 0, 1, or 2;

R⁹⁹ is selected from the group consisting of sulfur, SO, SO₂, NH,N-alkyl, oxygen, C═C (cis or trans), and C.tbd.C;

q is 0 or 1;

r is 0, 1, 2 or 3;

R¹² is NR¹³ R¹⁴, ##STR5## R¹³ -R¹⁶ are independently selected from thegroup consisting of H, hydroxy, amino, amidino, alkyl, cycloalkyl, acyl,aminoacyl, and phosphoryl and taken together (for example, one of R¹³ orR¹⁴ with one of R¹⁵ or R¹⁶ and the adjacent nitrogens and carbon formthe ring and the remaining R¹³ -R¹⁶ groups form the optionalsubstituent) may form an optionally substituted 5- or 6-membered ring;and

R¹⁷ is H or alkyl;

wherein alk₂ and R¹² taken together may form an optionally substituted 5or 6 member non-aromatic heterocycle.

Specific examples of heterocyclic groups known in the chemistry artsinclude the following (shown linked to a sulfur atom in each case, asabove): ##STR6##

Structures IV, V and VI show a vinyl group linking the cephem nucleus toan arylthio or heteroarylthio group. It is to be understood that eachcarbon atom of the vinyl group may optionally in addition be substitutedby a lower alkyl group, such as a methyl or ethyl group.

Preferred compounds include those compounds wherein R¹¹ is H or halogen,R¹² is NR¹³ R¹⁴, ##STR7## p is 0 or 1, q is 0 or 1, r is 1, 2 or 3, R¹³-R¹⁶ are H or lower alkyl, and R¹⁷ is H or lower alkyl. Especiallypreferred compounds include those compounds wherein R¹¹

is hydrogen, R¹² is NR¹³ R¹⁴, ##STR8## or, p is 1, q is 1, r is 1, 2, or3, R¹³ -R¹⁶ are hydrogen, and R¹⁷ is hydrogen or lower alkyl. There arepreferably one or two [(CH₂)_(p) (S)_(q) (CH₂)_(r) R¹² ] groups present.There are preferably three or four R¹¹ groups present.

Preferred compounds include those wherein:

(1) R¹ is NHC(0)ZR³

Z is --CH₂ (X)_(m) --

X is S

m is 1

(2) R¹ is NHC(0)ZR³

Z is --C(NOR⁶)--

R⁶ is selected from the group consisting of hydrogen, methyl,2-fluoroethyl, cyclopropylmethyl, allyl, dichloroallyl and cyclopentyl,and R³ is selected from the group consisting of phenyl, 2-thienyl,2-furyl, 2-aminothiazol-4-yl, 2-amino-5-chlorothiazol-4-yl, and2-aminothiadiazol-4-yl;

(3) R¹¹ is H or halogen

(4) R¹² is NR¹³ R¹⁴, ##STR9## (5) p is 0 or 1 q is 0 or 1 r is 1, 2 or3; and/or (6) R¹³ -R¹⁷ are H or lower alkyl.

Preferred compounds include those wherein alk₂ and R¹² taken together anoptionally substituted 5 or 6 member carbon ring containing a singlenitrogen. In especially preferred embodiments, alk₂ and R¹² takentogether form a substitutent (shown below on a sulfur atom) selectedfrom the group consisting of ##STR10##

Other preferred examples are shown below linked through a sulfur atom toa 4-pyridyl substituent: ##STR11##

Without being bound to any particular theory regarding operation of theinvention, it is noted that there are several novel structural featuresof the invention which are believed to contribute to the substantiallyimproved functional properties. In particular, chlorine substituents onheterocyclic R³ groups appear to impact 2-4 fold improved MIC valuescompared to the compounds with non-chlorinated heterocyclic R³ groups.

Preferred pharmaceutically acceptable salts include (1) inorganic saltssuch as chloride, bromide, iodide, nitrate, phosphate or sulfate; (2)carboxylate salts such as acetate, propionate, butyrate, maleate, orfumarate; (3) alkylsulfonates such as methanesulfonate, ethanesulfonate,2-hydroxyethylsulfonate, n-propylsulfonate or iso-propylsulfonate; and(4) hydroxycarboxylates such as lactate, malate, and citrate.

In another embodiment, the present invention provides for compositionscomprising an amount of a compound of Structure II, III, IV, V or VIeffective to treat bacterial infections in mammals arising from bacteriaresistant to β-lactam antibiotics.

In still another embodiment, the present invention includes methods fortreating a bacterial infection in a mammal arising from bacteriaresistant to β-lactam antibiotics comprising administering to a mammalsuffering from such an infection a therapeutically effective amount of acompound of Structure II, III, IV, V or VI. Of course, the compounds ofthe present invention also have utility in compositions and methods totreat mammals infected with bacteria that are sensitive to conventionalβ-lactam antibiotics.

In another embodiment, this invention features compounds and methods forpreparation of an intermediate VII, which is useful for preparingcompounds having particularly potent activity againstmethicillin-resistant Staphylococci and ampicillin-resistantEnterococci. ##STR12##

Other features and advantages of the invention will be apparent from thefollowing description of the preferred embodiments thereof and from theclaims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1-5 show preferred synthetic schemes for producing compounds ofthe present invention.

FIG. 1 shows preparation of the cephem.

FIG. 2 shows preparation of the C(7)-substituent.

FIG. 3 shows preparation of the 3-substituent.

FIG. 4 shows the final assembly.

FIG. 5 shows deprotection and salt formation.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

I. Definitions

As used herein, the term "alkyl" denotes branched or unbranchedhydrocarbon chains, preferably containing between one and six, morepreferably one and four, carbon atoms, such as, e.g., methyl, ethyl,n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, and2-methylpentyl. These groups may be optionally substituted with one ormore functional groups which are attached commonly to such chains, suchas, e.g., hydroxyl, bromo, fluoro, chloro, iodo, mercapto or thio,cyano, alkylthio, heterocycle, aryl, heteroaryl, carboxyl,alkoxycarbonyl, alkyl, alkenyl, nitro, amino, alkoxyl, amido, andoptionally substituted isothioureido, amidino, guanidino, and the liketo form alkyl groups such as trifluoromethyl, 3-hydroxyhexyl,2-carboxypropyl, 2-fluoroethyl, carboxymethyl, 4-cyanobutyl,2-guanidinoethyl, 3-N,N'-dimethylisothiouroniumpropyl, and the like.

The term "alkenyl" denotes an alkyl group as defined above having atleast one double bond, e.g., allyl, 3-hydroxy-2-buten-1-yl,1-methyl-2-propen-1-yl and the like.

The term "aryl" denotes a chain of carbon atoms an which form an leastone aromatic ring having preferably between about 6-14 carbon atoms,such as, e.g., phenyl, naphthyl, indenyl, and the like, and which may besubstituted with one or more functional groups which are attachedcommonly to such chains, such as, e.g., hydroxyl, bromo, fluoro, chloro,iodo, mercapto or thio, cyano, cyanoamido, alkylthio, heterocycle, aryl,heteroaryl, carboxyl, alkoxycarbonyl, alkyl, alkenyl, nitro, amino,alkoxyl, amido, and the like to form aryl groups such as biphenyl,iodobiphenyl, methoxybiphenyl, anthryl, bromophenyl, iodophenyl,chlorophenyl, hydroxyphenyl, methoxyphenyl, formylphenyl, acetylphenyl,trifluoromethylthiophenyl, trifluoromethoxyphenyl, alkylthiophenyl,trialkylammoniumphenyl, amidophenyl, thiazolylphenyl, oxazolylphenyl,imidazolylphenyl, imidazolylmethylphenyl, cyanophenyl, pyridylphenyl,pyrrolylphenyl, pyrazolylphenyl, triazolylphenyl, tetrazolylphenyl andthe like.

The term "heterocycle" denotes a chain of carbon and at least onenon-carbon atoms which together form one or more aromatic ornon-aromatic rings having preferably between about 5-14 atoms, such as,e.g., furyl, thienyl, imidazolyl, indolyl, pyridinyl, thiadiazolyl,thiazolyl, piperazinyl, dibenzfuranyl, dibenzthienyl. These rings may beoptionally substituted with one or more functional groups which areattached commonly to such rings, such as, e.g., hydroxyl, bromo, fluoro,chloro, iodo, mercapto or thio, cyano, cyanoamido, alkylthio,heterocycle, aryl, heteroaryl, carboxyl, oxo, alkoxycarbonyl, alkyl,alkenyl, nitro, amino, alkoxyl, amido, and the like to form rings suchas, e.g., 2-aminothiazol-4-yl, 2-amino-5-chlorothiazol-4-yl,2-amino-thiadiazol-4-yl, 2,3-dioxopiperazinyl, 4-alkylpiperazinyl,2-iodo-3-dibenzfuranyl and 3-hydroxy-4-dibenzthienyl and the like.

The term "heteroaromatic" or "heteroaryl" (HetAr) denotes an aromaticheterocycle as defined above.

The term "heterotricycle" denotes an aromatic heterocyclic substituentas defined above which comprises three aromatic rings.

The term "heterocyclecarbonyl" denotes the group --C(O)Het, where Het isheterocycle as defined above.

The term "alkoxyl" denotes the group --OR, where R is alkyl as definedabove, such as methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy,sec-butoxy, iso-butoxy, tert-butoxy, trifluoromethoxy,3-hydroxyhexyloxy, 2-carboxypropyloxy, 2-fluoroethoxy, carboxymethoxyand cyanobutyloxy and the like.

The term "alkylthio" denotes the group --SR, where R is alkyl as definedabove, such as methylthio, ethylthio, n-propylthio, iso-propylthio,n-butylthio, sec-butylthio, iso-butylthio, tert-butylthio,trifluoromethylthio, 3-hydroxyhexylthio, 2-carboxypropylthio,2-fluoroethylthio, carboxymethylthio and cyanobutylthio and the like.

The term "acyl" denotes groups --C(O)R, where R is alkyl as definedabove, such as formyl, acetyl, propionyl, or butyryl.

The term "aryloxy" denotes groups --OAr, where Ar is an aryl group asdefined above.

The term "aralkyl" denotes groups --RAr, where R is alkyl and Ar isaryl, both as defined above.

The term "heteroaralkyl" denotes groups --RHetAr where R is alkyl andHetAr is heteroaryl as defined above.

The term "trialkylsilyl" denotes the group RR'R"Si--, where R, R' and R"are alkyl as defined above.

The term "trialkylammonium" denotes the group [RR'R"N--]⁺, where R, R'and R" are alkyl as defined above.

The term "amino" denotes the group NRR', where R and R' mayindependently be alkyl, aryl or acyl as defined above, or hydrogen.

The term "carboxamido" denotes the group --C(O)NRR', where R and R' mayindependently be alkyl, aryl or acyl as defined above, or hydrogen.

The term "cyanoamido" refers to the group --NH--C.tbd.N.

The term "β-lactam resistant bacteria" refers to bacteria against whicha β-lactam antibiotic has an minimum inhibitory concentration (MIC)greater than 32 mg/ml.

The term "methicillin-resistant bacteria" refers to bacteria that areresistant to methicillin. Examples of such bacteria are provided inTable 1 and are identified Meth^(R). The term "methicillin sensitivebacteria" refers to bacteria that are sensitive to methicillin. Examplesof such bacteria are provided in Table 1 and are identified Meth^(S).

The term "prodrug" refers to an agent that is converted into the parentdrug in vivo. Prodrugs may be easier to administer than the parent drugin some situations. For example, the prodrug may be bioavailable by oraladministration but the parent is not, or the prodrug may improvesolubility to allow for intravenous administration.

II. Compounds of the Invention

The present invention provides compounds, methods and compositionseffective to treat bacterial infections, and, especially, infectionsarising from bacteria which have developed resistance to conventionalβ-lactam antibiotics. More importantly, the present invention providescompounds, methods and compositions effective to treat bacterialinfections arising from bacteria which have developed resistance toconventional cephalosporin antibiotics.

A. Synthesis of Compounds of Structure II

The compounds of the present invention may be readily prepared inaccordance with the following schemes. However, it will be appreciatedthat other synthetic pathways for forming the compounds of the inventionare available and that the following is offered merely by way ofexample, and not limitation. It will be further recognized that variousprotecting and deprotecting strategies will be employed which arestandard in the art (see, e.g., Green and Wuts). Those of skill in theart will recognize that the selection of any particular protecting group(e.g., a carboxy protecting group) will depend on the stability of theprotected moiety with respect to subsequent reaction conditions.

Generally, the synthesis of the cephalosporins of the present inventionmay be achieved using well-known methods and readily available materials(see, e.g., March; Larock, COMPREHENSIVE ORGANIC TRANSFORMATIONS (VCHPublishers, 1989) ; and G. I. Georg, THE ORGANIC CHEMISTRY OF β-LACTAMS,(VCH 1992), each of which is incorporated herein by reference). As shownbelow in Scheme 1, treatment of the cephem triflate 1 with the desiredoptionally protected thiolate nucleophile 2, using standard methods suchas those described in Farina et al., J. Org. Chem, 54: 4962 (1989) andU.S. Pat. No. 4,870,168 to Baker, et al., (both of which areincorporated herein by reference), provides the 3-thio derivative 3.Subsequent deprotection using procedures known to those skilled in theart affords the biologically active 4-carboxycephem 4. ##STR13##

Compound 1 is formed readily from commercially available startingmaterials, such as the reaction of(7R)-7-[(phenylacetyl)amino]-3-hydroxy-3-cephem-4-carbox ylic acid(Otsuka Chemical Co., Ltd., Otsuka, Japan) with triflic anhydride(Aldrich, Milwaukee, Wis.), using known procedures (see. e.g., Farina;and U.S. Pat. No. 4,870,168 to Baker, et al.) Other 3-hydroxy-3-cephemsmay be formed from the ozonolysis of 3-exomethylene cephems using knownprocedures (see. e.g., Farina). Similarly, the thiolate nucleophile 2may be formed using known procedures and commercially available startingmaterials.

The substituent R¹ may be any of the groups described above and areeither available commercially (e.g., from Aldrich, Milwaukee, Wis.) orcan be formed using known techniques and starting materials (see, e.g.,March; Larock). These groups can be substituted for those present on thestarting material by variety of well known techniques (see. e.g.,Barrett, J. C. S. Perkin I, 1629 (1979) or Chauvette, J. Org. Chem. 36:1259 (1971), both of which are incorporated herein by reference), suchas by transamination of an existing substituent for the desiredsubstituent, or hydrolytic removal of the existing substituent followedby reaction with a suitably reactive form of desired substituent, suchas an acyl chloride. Again, the appropriate reagents and techniques willbe apparent to those of skill in the art.

The carboxyl group R² may be those protecting groups amenable toreductive cleavage, such as benzyl, p- or o-nitrobenzyl,2,2,2-trichloroethyl, allyl, cinnamyl, benzhydryl, 2-chloroallyl and thelike. Alternatively, R² may be a protecting group amenable to acidiccleavage, such as t-butyl, t-amyl, trityl, 4-methoxytrityl,4,4'-dimethoxytrityl, trimethylsilyl, t-butyldimethylsilyl, phenacyl,β-(trimethylsilyl)ethyl, benzyl, 4-(or 2-methoxybenzyl,2,4-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl,methoxymethyl, benzhydryl, or 3,3-dimethylallyl. Preferred protectinggroups are p-methoxybenzyl, p-nitrobenzyl, allyl and benzhydryl. Suchgroups may be attached to the unprotected carboxyl group of thecephalosporin starting material using known reagents and techniques,such as those described in Green and Wuts.

B. Synthesis of Compounds of Structure III

The compounds of general structure III are prepared similarly to thoseof general structure II. In most cases, a key step is the coupling of asubstituted heteroarylthiolate with cephem triflate 1 or a functionallyequivalent cephem having an alternative leaving group at C-(3).Compounds of structure III where the ring containing G, H, J, L and M is4-pyridyl may also be prepared as exemplified in FIG. 4.

C. Synthesis of Compounds of Structures IV, V and VI

The compounds of general structure IV, V and VI are prepared by couplingan aromatic or heteroaromatic thiolate with tosylvinyl cephem sulfoxide6. Compound 6 is formed readily from commercially available startingmaterials using known procedures (see. e.g., Farge et al., U.S. Pat. No.4,307,116). The requisite aromatic or heteroaromatic thiols are preparedby a variety of methods known in the literature, as described in theExamples. As shown below in Scheme 2, treatment of the cephemintermediate 6 with a desired optionally protected thiolate nucleophile,such as 5, using methods such as those described in Farge et al., U.S.Pat. No. 4,307,116 (which is incorporated herein by reference), providesthe 3-thiovinyl derivative 7. Sulfoxide reduction and 5 subsequentdeprotection using procedures known to those skilled in the art affordsthe biologically active 4-carboxycephem 8. ##STR14## D. Synthesis ofCompounds of Structure VII

A preferred synthetic route for preparation of these intermediates isset forth in FIG. 2. In particular, the step of converting 6 to 7 shownin FIG. 2 and described in Example 18 is especially useful in providingcompounds having substantially improved activity againstmethicillin-resistant and ampicillin-resistant bacteria.

III. Pharmaceutical Applications and Preparations

According to this invention, a therapeutically or pharmaceuticallyeffective amount of a cephalosporin and particularly, a compound ofStructure II, III, IV, V or VI, is administered to a mammal sufferingfrom an methicillin-resistant bacterial infection (or other β-lactamresistant bacterial infections, such as vancomycin-resistant orampicillin-resistant infections), especially resistant S. aureus, in anamount effective to at least partially relieve the infection. Especiallyimportant are infections resulting from strains having similar activityto strains such as S. aureus Col (Meth^(R)) (lac⁻), S. aureus 76(Meth^(R)) (lac⁺), E. f.ae butted.cium ATCC 35667, or E. f.aebutted.calis ATCC 29212. Again, such compounds are also effectiveagainst bacteria sensitive to methicillin, vancomycin, and/or ampicillinand therefore have utility in such compositions and methods.

The compositions containing the compound(s) of the invention can beadministered for prophylactic and/or therapeutic treatments. Intherapeutic applications, the compositions are administered to a patientalready suffering from an infection, as described above, in an amountsufficient to cure or at least partially arrest the symptoms of theinfection. An amount adequate to accomplish this is defined as"therapeutically effective amount or dose." Amounts effective for thisuse will depend on the severity and course of the infection, previoustherapy, the patient's health status and response to the drugs, and thejudgment of the treating physician. In prophylactic applications,compositions containing the compounds of the invention are administeredto a patient susceptible to or otherwise at risk of a particularinfection. Such an amount is defined to be a "prophylactically effectiveamount or dose." In this use, the precise amounts again depend on thepatient's state of health, weight, and the like.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, can be reduced, as a function ofthe symptoms, to a level at which the improved condition is retained.When the symptoms have been alleviated to the desired level, treatmentcan cease. Patients can, however, require intermittent treatment on along-term basis upon any recurrence of the disease symptoms.

In general, a suitable effective dose of the compound of the inventionwill be in the range of 0.1 to 1000 milligram (mg) per recipient perday, preferably in the range of 1 to 100 mg per day. The desired dosageis preferably presented in one, two, three, four or more subdosesadministered at appropriate intervals throughout the day. These subdosescan be administered as unit dosage forms, for example, containing 5 to1000 mg, preferably 10 to 100 mg of active ingredient per unit dosageform. Preferably, the compounds of the invention will be administered inamounts of between about 2.0 mg/kg to 250 mg/kg of patient body weight,between about one to four times per day.

While it is possible to administer the active ingredient of thisinvention alone, it is preferable to present it as part of apharmaceutical formulation. The formulations of the present inventioncomprise at least one compound or inhibitor of this invention in atherapeutically or pharmaceutically effective dose together with one ormore pharmaceutically or therapeutically acceptable carriers. Solidcarriers inlcude, e.g., starch, lactose, dicalcium phosphate,microcrystalline cellulose, sucrose, and kaolin, and optionally othertherapeutic ingredients. Liquid carriers include, e.g., sterile water,polyethylene glycols, non-ionic surfactants, and edible oils such ascorn, peanut and sesame oils. In addition, various adjuvants such as arecommonly used in the art may be included. For example: flavoring agents,coloring agents, preservatives, and antioxidants, e.g., vitamin E,ascorbic acid, BHT and BHA. Various other considerations are described,e.g., in Gilman et al. (eds) (1990) Goodman and Gilman's: ThePharmacological Basis of Therapeutics, 8th Ed., Pergamon Press; andRemington's supra. Methods for administration are discussed therein,e.g., for oral, intravenous, intraperitoneal, or intramuscularadministration, and others. Pharmaceutically acceptable carriers willinclude water, saline, buffers, and other compounds described, e.g., inthe MERCK INDEX, Merck & Co., Rahway, N.J. Generally, preferred routesof administration are intravenous and intraperitoneal.

These pharmacological agents can be in a variety of forms. Theseinclude, for example, solid, semi-solid and liquid dosage forms, such astablets, pills, powders, liquid solutions or suspensions, liposomes,injectable and infusible solutions. The preferred form depends on theintended mode of administration and therapeutic application. Generally,a pharmacologically acceptable salt of the compound will be used tosimplify preparation of the composition. Preferred salts include sodium,potassium, arginine, glycine, alanine, threonine. These are prepared,preferably, in water suitably mixed with a surfactant such ashydroxypropylcellulose.

Depending on the specific conditions being treated, such agents may beformulated and administered systemically or locally. Techniques forformulation and administration may be found in Remington'sPharmaceutical Sciences, 18th ed., Mack Publishing Co., Easton, Pa.(1990). Suitable routes may include oral, rectal, transdermal, vaginal,transmucosal, or intestinal administration; parenteral delivery,including intramuscular subcutaneous, intramedullary injections, as wellan intrathecal, direct intraventricular, intravenous, intraperitoneal,intranasal, or intraocular injections, just to name a few.

For injection, the agents of the invention may be formulated in aqueoussolutions, preferably in physiologically compatible buffers such asHanks' solution, Ringer's solution, or physiological saline buffer. Forsuch transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants aregenerally known in the art.

In soft capsules, the active compounds may be dissolved or suspended insuitable liquids, such as fatty oils, liquid paraffin, or liquidpolyethylene glycols. In addition, stabilizers may be added.

IV. Biological Activity

In Vitro Antibacterial Evaluation

The compounds of the invention were evaluated against several β-lactamresistant (for example methicillin-resistant, vancomycin resistant,and/or ampicillin-resistant) bacteria strains by determining the minimuminhibitory concentration (MIC, μg/ml) of each compound with respect toeach strain. The MIC, the lowest concentration of antibiotic whichinhibits growth of the test organism, was determined by the agardilution method.

To determine the MIC for bacterial isolates, the test compound wasincorporated in a series of two-fold dilutions into liquifiedMueller-Hinton agar. Upon solidification, a number of differentbacterial strains were spot inoculated with a replicating device ontothe agar surface. After overnight incubation, the MIC breakpoint wasdetermined as the lowest drug concentration that completely inhibitedgrowth, disregarding a single colony or a faint haze. The proceduresused in these stuides have been standardized by the National Committeefor Clinical Laboratory Standards (NCCLS), as per the NCCLS publicationentitled METHODS FOR DILUTION ANTIMICROBIAL SUSCEPTIBILITY TESTS (1991),which is incorporated herein by reference.

Aliquots of antimicrobial agents were prepared in phosphate bufferedsaline (PBS) at pH 7.2. Tween 20 or DMSO was used as a solubilizingvehicle as needed. Standard methods of vortexing, sonicating and gentleheat were used to facilitate solubilizing the test agent. Typically, theconcentration of the stock solution was 10× that of the highest drugconcentration tested. A 1.28 mg/mL stock solution was used with asubsequent highest working concentration of 128 μg/mL. Serial two-folddilutions were done through ≦0.25 μg/mL. Each drug level was tested induplicate. Two-fold drug dilutions were done in sterile 50 mL tubes witha final drug volume of 5 mL. Upon the addition of 45 mL of molten agar,a 10-fold dilution resulted. Two, 25 mL. plates were then poured into15×150 mm square Petri plates with grids and allowed to harden.

A control plate with a reference drug, either cefotaxime, vancomycin orimipenem, was used as the positive growth control. Stock concentrationsof reference antibiotics were prepared and frozen at -80° C. Uponpreparation, the control plates were sealed and stored in therefrigerator for up to 1 week prior to use; however, imipenem controlplates had to be prepared just prior to use. All test plates were usedwithin 24 hours of preparation.

Satisfactory results were obtained where the inoculum contained about10⁴ colony forming units (cfu)±0.5 logs. Starting with pure cultures ofthe test isolates on agar plates, a few isolated colonies weretransferred to a tube of nutrient broth and allowed to grow 4-6 hours at35-36° C. to reach log-phase growth. Dropwise addition of the brothculture to PBS was done to match a 0.5 McFarland turbidity standardequal to 10⁸ cfu/mL. This was further diluted ten-fold in PBS to reach aworking inoculum concentration of 10⁷ cfu/mL. When 1 μL of the workinginoculum was applied to the agar surface a concentration of about 10⁴cfu per spot was obtained.

Disposable sterile 1 μL loops were used to inoculate test plates, witheach isolate in a designated grid on the agar plate. An alternate methodof inoculation involved the use of a replica plater, a device with 48steel pins allowing the simultaneous inoculation of multiple isolates.After the spots had dried, the plates were incubated at 35-36° C. for16-20 hours. Endpoints were assessed as the minimum inhibitoryconcentration (MIC) of antimicrobial agent.

The novel agents of this invention are notable for their enhancedactivity against S. aureus Col and Enterococci (E. faecium and E.faecalis). The S. aureus Col strain is a high-level PBP2a producer,whereas S. aureus Col 8A, its isogenic partner, lacks PBP2a.

Certain compounds show broad activity against both S. aureus Col and S.aureus Col 8A, as well as Enterococci. The S. aureus Col 8A strain washighly responsive to all test agents including the Cefotaxime control.Thus, the compounds of the present invention are effective againstPBP2a-producing bacteria. Certain compounds show potent activity againstenterococci. Certain other compounds of the present invention, areeffective against E. coli in addition to Gram-positive organisms.

In Vivo Antibacterial Evaluation

Compounds with superior activity in vitro when compared to referenceantibiotics, are further evaluated in a murine model for lethalbacteremic peritonitis.

Groups of 5 female Swiss-Webster mice (Simonsen, Gilroy, Calif.) eachare challenged by the intraperitoneal (IP) route with tenfold incrementsof a bacterial inoculum. This permits calculation of the mean lethaldose (LD₅₀) and the LD₁₀₀. For preliminary evaluation of a newantibiotic, mice are challenged IP with an LD₁₀₀ titer of bacteria. Intwo equal doses administered at the time of bacterial challenge and 2hours later, groups of 10 mice each are treated subcutaneously withtwo-fold increments of the test drug and an antibiotic of known efficacyin mice and humans (i.e., positive control). Mice are observed for 72 h.Those alive at 72 h are considered long term survivors. The total drugdose in mg/kg that protects 50% of mice in a group from death is termedthe mean protective dose (PD₅₀) . PD₅₀ s are similarly determined forseveral pathogens. The quantitative endpoints for the new drug are thencompared with those obtained with reference antibiotics.

Six ten-fold dilutions of inoculum suspended in 0.5 mL of sterilized 7%hog gastric mucin (Sigma) are injected IP in groups of 5 mice each. Acontrol group of 5 mice receive mucin alone. Mice are observed for 72 h.Those alive at 72 h are considered long term survivors. The mean lethaldose (LD₅₀) and 100% lethal dose (LD₁₀₀) are determined by the probittest.

For antibiotic efficacy studies, mice are challenged IP with bacterialtiters that will afford an LD₁₀₀ for the test strain. In two equal dosesadministered at the time of bacterial challenge and 2 hours later,groups of 10 mice each are treated by the subcutaneous route (SC) withtwofold increments of the test antibiotic; another group is treatedsimilarly with a reference antibiotic of known efficacy in animals andman. Drug doses can range from 0.01 to 512 mg/kg. If the drug is poorlysoluble, Tween 20 or propylene glycol will be employed to solubilize thedrug. Animals are observed for 72 h. The 50% protective dose (PD₅₀) iscalculated in mg/kg by the probit method. The PD₅₀ is the same as the50% effective dose (ED₅₀) and the 50% curative dose (CD₅₀). Samples ofblood from the hearts of all animals that die and from half the micethat survive are cultured on brain-heart infusion agar. Animals thatreceived a protective dosage of the test drug will be alive at 72 h,although they may appear moderately ill to very ill during theobservation period. Infected, placebo-treated control mice and thosereceiving non-effective i.e. lower dosages of the test drug willdemonstrate a high rate of mortality. Most of these mice will die within6 to 48 h. Those alive at 72 h will be considered long term survivors.

                                      TABLE 1                                     __________________________________________________________________________    Antimicrobial Properties of 7-(Acylamido)-3-(arylthio)cephems                 __________________________________________________________________________    Organism      Imipenem                                                                           1   2    3   4   5    6   7    8   9   10                  __________________________________________________________________________    S. aureus ATCC 29213                                                                        ≦0.25                                                                       0.13                                                                              ≦0.06                                                                       0.5 0.25                                                                              0.13 0.25                                                                              0.5  1   0.25                                                                              0.5                 S. aureus Col8A (Meth.sup.S)(lac-)                                                          ≦0.25                                                                       ≦0.06                                                                      ≦0.06                                                                       0.5 0.25                                                                              ≦0.06                                                                       0.13                                                                              0.5  1   0.13                                                                              0.5                 S. aureus PC1 (Meth.sup.S)(lac+)                                                            ≦0.25                                                                       1   0.5  0.5 0.25                                                                              0.25 0.5 2    1   1   0.5                 S. aureus ATCC 13709 (Meth.sup.S)                                                           ≦0.25                                                                       --  --   0.5 0.25                                                                              ≦0.06                                                                       ≦0.06                                                                      0.5  1   ≦0.06                                                                      0.5                 S. aureus Col (Meth.sup.R)(lac-)                                                            32   4   1    8   8   8    2   8    2   1   2                   S. aureus 76 (Meth.sup.R)(lac+)                                                             32   8   4    8   8   16   8   8    4   4   2                   S. aureus ATCC 33593 (Meth.sup.R)                                                           32   8   4    8   4   16   8   8    4   4   2                   S. aureus Spain #356 (Meth.sup.R)                                                           32   8   4    8   8   16   8   8    4   4   2                   S. haemolyticus 05 (Meth.sup.R)                                                             64   16  4    8   8   16   16  32   4   16  4                   E. faecalis ATCC 29212                                                                      ≦0.25                                                                       1   0.5  0.5 0.5 2    0.5 1    0.5 1   0.5                 E. faecium ATCC 35667                                                                       4    2   1    2   2   4    2   4    2   1   2                   E. faecium VanA (Van.sup.R)                                                                 4    8   4    8   4   32   8   16   8   4   4                   E. faecalis VanB (Van.sup.R)                                                                0.5  4   0.5  1   4   4    8   2    0.5 2   1                   E. faecium A491 (Amp.sup.R)                                                                 >128 >32 >32  >32 >32 >32  >32 >32  32  >32 >32                 E. coli ATCC25992                                                                           ≦0.25                                                                       >32 16   2   4   >32  >32 4    2   >32 4                   P. aeruginosa ATCC 27853                                                                    1    >32 >32  >32 >32 >32  >32 >32  >32 >32 >32                 __________________________________________________________________________    Organism      Imipenem                                                                           11  12   13  14  15   16  17   18  19  20                  __________________________________________________________________________    S. aureus ATCC 29213                                                                        ≦0.25                                                                       0.5 0.5  2   0.5 1    0.25                                                                              0.25 1   0.5 0.5                 S. aureus Col8A (Meth.sup.S)(lac-)                                                          ≦0.25                                                                       1   0.5  2   0.5 1    0.25                                                                              0.5  1   0.5 0.5                 S. aureus PC1 (Meth.sup.S)(lac+)                                                            ≦0.25                                                                       1   0.5  2   0.5 1    0.25                                                                              0.25 1   0.5 0.5                 S. aureus ATCC 13709 (Meth.sup.S)                                                           ≦0.25                                                                       0.5 0.25 2   0.25                                                                              0.5  0.13                                                                              0.25 1   0.5 0.5                 S. aureus Col (Meth.sup.R)(lac-)                                                            32   4   2    4   4   4    0.5 2    44  8   2                   S. aureus 76 (Meth.sup.R)(lac+)                                                             32   8   2    8   4   4    1   4    8   8   4                   S. aureus ATCC 33593 (Meth.sup.R)                                                           32   4   2    8   2   4    1   2    4   8   2                   S. aureus Spain #356 (Meth.sup.R)                                                           32   8   2    8   4   4    1   4    8   8   4                   S. haemolyticus 05 (Meth.sup.R)                                                             64   4   2    8   4   8    2   8    8   16  4                   E. faecalis ATCC 29212                                                                      ≦0.25                                                                       0.25                                                                              0.25 1   0.25                                                                              2    ≦0.06                                                                      0.13 0.25                                                                              1   1                   E. faecium ATCC 35667                                                                       4    2   1    4   1   2    0.5 1    2   1   1                   E. faecium VanA (Van.sup.R)                                                                 4    4   4    8   2   4    0.25                                                                              0.25 2   2   2                   E. faecalis VanB (Van.sup.R)                                                                0.5  0.5 0.5  2   0.25                                                                              0.5  0.13                                                                              0.5  0.5 0.5 0.5                 E. faecium A491 (Amp.sup.R)                                                                 >128 >32 >32  >32 >32 >32  8   >32  >32 >32 >32                 E. coli ATCC25992                                                                           ≦0.25                                                                       8   8    8   4   8    8   2    8   8   4                   P. aeruginosa ATCC 27853                                                                    1    >32 >32  >32 >32 >32  >32 >32  >32 >32 >32                 __________________________________________________________________________    Organism      Imipenem                                                                           21  22   23  24  25   26  27   28  29  30                  __________________________________________________________________________    S. aureus ATCC 29213                                                                        ≦0.25                                                                       0.25                                                                              0.25 1   1   0.25 0.13                                                                              0.25 1   0.25                                                                              0.25                S. aureus Col8A (Meth.sup.S)(lac-)                                                          ≦0.25                                                                       0.25                                                                              0.5  0.5 1   0.25 0.13                                                                              0.13 0.25                                                                              0.25                                                                              0.13                S. aureus PC1 (Meth.sup.S)(lac+)                                                            ≦0.25                                                                       0.5 1    0.5 1   0.25 0.13                                                                              0.25 0.5 0.25                                                                              ≦0.25        S. aureus ATCC 13709 (Meth.sup.S)                                                           ≦0.25                                                                       1   1    0.5 1   0.25 ≦0.06                                                                      0.13 0.13                                                                              0.13                                                                              ≦0.25        S. aureus Col (Meth.sup.R)(lac-)                                                            32   1   2    2   8   1    0.5 0.5  2   1   0.5                 S. aureus 76 (Meth.sup.R)(lac+)                                                             32   1   2    4   8   2    1   1    2   1   2                   S. aureus ATCC 33593 (Meth.sup.R)                                                           32   1   2    4   4   1    1   1    4   2   1                   S. aureus Spain #356 (Meth.sup.R)                                                           32   1   2    4   4   2    1   1    2   2   1                   S. haemolyticus 05 (Meth.sup.R)                                                             64   2   2    4   8   2    2   2    4   2   1                   E. faecalis ATCC 29212                                                                      ≦0.25                                                                       0.13                                                                              0.13 0.5 0.25                                                                              0.13 ≦0.06                                                                      ≦0.06                                                                       0.13                                                                              ≦0.06                                                                      ≦0.06        E. faecium ATCC 35667                                                                       4    0.5 1    2   2   1    0.25                                                                              0.25 1   0.25                                                                              0.5                 E. faecium VanA (Van.sup.R)                                                                 4    1   2    4   4   2    0.5 0.5  4   0.5 1                   E. faecalis VanB (Van.sup.R)                                                                0.5  0.13                                                                              0.5  1   0.5 0.13 0.13                                                                              ≦0.06                                                                       0.25                                                                              ≦0.06                                                                      ≦0.06        E. faecium A491 (Amp.sup.R)                                                                 >128 8   16   >32 >32 32   8   4    16  4   8                   E. coli ATCC25992                                                                           ≦0.25                                                                       32  8    2   8   4    >32 8    2   8   8                   P. aeruginosa ATCC 27853                                                                    1    >32 >32  >32 >32 >32  >32 32   32  >32 >32                 __________________________________________________________________________    Organism      Imipenem                                                                           31  32   33  34  35   36  37   38  39  40                  __________________________________________________________________________    S. aureus ATCC 29213                                                                        ≦0.25                                                                       0.25                                                                              0.25 0.25                                                                              ≦0.06                                                                      0.25 0.5 0.5  0.13                                                                              0.25                                                                              0.25                S. aureus Col8A (Meth.sup.S)(lac-)                                                          ≦0.25                                                                       0.25                                                                              0.25 0.25                                                                              ≦0.06                                                                      0.25 0.25                                                                              ≦0.06                                                                       0.13                                                                              0.5 0.5                 S. aureus PC1 (Meth.sup.S)(lac+)                                                            ≦0.25                                                                       0.25                                                                              0.5  0.25                                                                              0.13                                                                              0.5  0.25                                                                              1    0.25                                                                              0.5 0.13                S. aureus ATCC 13709 (Meth.sup.S)                                                           ≦0.25                                                                       0.25                                                                              0.13 0.13                                                                              ≦0.06                                                                      0.25 0.25                                                                              ≦0.06                                                                       0.13                                                                              0.25                                                                              0.25                S. aureus Col (Meth.sup.R)(lac-)                                                            32   2   1    1   1   2    1   0.5  1   2   2                   S. aureus 76 (Meth.sup.R)(lac+)                                                             32   2   2    1   1   2    2   4    1   4   2                   S. aureus ATCC 33593 (Meth.sup.R)                                                           32   2   2    2   1   2    1   16   1   4   2                   S. aureus Spain #356 (Meth.sup.R)                                                           32   2   2    1   1   2    1   8    1   4   2                   S. haemolyticus 05 (Meth.sup.R)                                                             64   4   4    2   2   4    2   8    2   8   4                   E. faecalis ATCC 29212                                                                      ≦0.25                                                                       0.5 ≦0.06                                                                       0.13                                                                              ≦0.06                                                                      ≦0.06                                                                       0.25                                                                              0.25 0.13                                                                              0.25                                                                              0.13                E. faecium ATCC 35667                                                                       4    0.5 0.5  0.5 0.25                                                                              0.25 0.5 0.5  1   1   1                   E. faecium VanA (Van.sup.R)                                                                 4    1   1    2   0.5 0.5  1   2    2   2   2                   E. faecalis VanB (Van.sup.R)                                                                0.5  0.25                                                                              0.13 0.13                                                                              ≦0.06                                                                      0.25 0.13                                                                              0.5  0.13                                                                              0.25                                                                              0.25                E. faecium A491 (Amp.sup.R)                                                                 >128 8   8    8   4   4    8   >32  >32 32  16                  E. coli ATCC25992                                                                           ≦0.25                                                                       8   8    1   4   8    16  >32  2   8   8                   P. aeruginosa ATCC 27853                                                                    1    >32 >32  32  >32 >32  >32 >32  >32 >32 >32                 __________________________________________________________________________    Organism      Imipenem                                                                           41  42   43  44  45   46  47   48  49  50                  __________________________________________________________________________    S. aureus ATCC 29213                                                                        ≦0.25                                                                       0.13                                                                              0.13 0.5 0.5 1    1   1    0.5 0.25                                                                              0.13                S. aureus Col8A (Meth.sup.S)(lac-)                                                          ≦0.25                                                                       0.25                                                                              0.25 0.5 0.5 1    0.5 0.5  0.5 0.5 0.25                S. aureus PC1 (Meth.sup.S)(lac+)                                                            ≦0.25                                                                       0.25                                                                              0.13 1   2   2    2   2    0.5 0.5 0.25                S. aureus ATCC 13709 (Meth.sup.S)                                                           ≦0.25                                                                       0.13                                                                              0.13 0.5 2   2    2   2    0.25                                                                              0.25                                                                              0.13                S. aureus Col (Meth.sup.R)(lac-)                                                            32   1   0.5  2   4   4    2   2    4   2   2                   S. aureus 76 (Meth.sup.R)(lac+)                                                             32   1   1    2   4   4    4   2    8   2   2                   S. aureus ATCC 33593 (Meth.sup.R)                                                           32   1   1    2   4   4    2   2    4   2   2                   S. aureus Spain #356 (Meth.sup.R)                                                           32   1   1    2   4   4    4   2    8   2   2                   S. haemolyticus 05 (Meth.sup.R)                                                             64   1   2    4   8   8    8   4    16  4   4                   E. faecalis ATCC 29212                                                                      ≦0.25                                                                       ≦0.06                                                                      ≦0.06                                                                       0.13                                                                              0.13                                                                              0.13 0.13                                                                              0.13 0.25                                                                              0.13                                                                              0.13                E. faecium ATCC 35667                                                                       4    0.25                                                                              0.25 1   4   2    2   2    1   1   1                   E. faecium VanA (Van.sup.R)                                                                 4    1   0.5  1   2   4    2   2    4   2   2                   E. faecalis VanB (Van.sup.R)                                                                0.5  ≦0.06                                                                      ≦0.06                                                                       0.25                                                                              --  --   --  --   0.5 0.13                                                                              0.5                 E. faecium A491 (Amp.sup.R)                                                                 >128 8   4    8   16  16   16  16   32  16  16                  E. coli ATCC25992                                                                           ≦0.25                                                                       4   2    8   8   16   8   8    4   8   4                   P. aeruginosa ATCC 27853                                                                    1    32  32   >32 >32 >32  >32 >32  >32 >32 >32                 __________________________________________________________________________    Organism      Imipenem                                                                           51  52   53  54  55   56  57   58  59  60                  __________________________________________________________________________    S. aureus ATCC 29213                                                                        ≦0.25                                                                       0.25                                                                              0.5  0.5 0.25                                                                              0.5  0.5 1    0.5 0.5 0.25                S. aureus Col8A (Meth.sup.S)(lac-)                                                          ≦0.25                                                                       0.5 0.25 0.5 0.25                                                                              0.5  0.5 1    0.5 0.25                                                                              0.25                S. aureus PC1 (Meth.sup.S)(lac+)                                                            ≦0.25                                                                       0.5 0.5  1   0.25                                                                              1    0.5 1    1   0.5 0.25                S. aureus ATCC 13709 (Meth.sup.S)                                                           ≦0.25                                                                       0.5 0.5  0.25                                                                              0.25                                                                              0.25 0.25                                                                              0.5  0.5 0.13                                                                              0.13                S. aureus Col (Meth.sup.R)(lac-)                                                            32   2   2    4   2   4    2   4    4   2   2                   S. aureus 76 (Meth.sup.R)(lac+)                                                             32   4   2    4   2   4    2   4    4   2   1                   S. aureus ATCC 33593 (Meth.sup.R)                                                           32   2   2    2   1   2    2   4    2   2   2                   S. aureus Spain #356 (Meth.sup.R)                                                           32   2   2    4   2   4    2   4    4   4   2                   S. haemolyticus 05 (Meth.sup.R)                                                             64   2   2    4   2   4    4   8    4   4   4                   E. faecalis ATCC 29212                                                                      ≦0.25                                                                       0.13                                                                              ≦0.06                                                                       0.13                                                                              0.13                                                                              0.25 0.13                                                                              0.13 0.5 0.5 0.5                 E. faecium ATCC 35667                                                                       4    0.13                                                                              ≦0.06                                                                       1   0.5 0.5  1   1    1   1   1                   E. faecium VanA (Van.sup.R)                                                                 4    1   1    2   1   2    1   2    2   2   2                   E. faecalis VanB (Van.sup.R)                                                                0.5  0.25                                                                              0.13 0.25                                                                              0.25                                                                              0.5  2   4    0.5 0.5 0.25                E. faecium A491 (Amp.sup.R)                                                                 >128 8   4    16  8   16   8   16   16  16  16                  E. coli ATCC25992                                                                           ≦0.25                                                                       8   2    4   8   8    8   4    8   8   8                   P. aeruginosa ATCC 27853                                                                    1    >32 >32  >32 >32 >32  >32 >32  >32 >32 >32                 __________________________________________________________________________    Organism      Imipenem                                                                           61  62   63  64  65   66  67   68  69                      __________________________________________________________________________    S. aureus ATCC 29213                                                                        ≦0.25                                                                       1   0.5  0.25                                                                              0.25                                                                              0.5  0.13                                                                              0.13 0.13                                                                              0.13                    S. aureus Col8A (Meth.sup.S)(lac-)                                                          ≦0.25                                                                       1   0.5  0.13                                                                              0.25                                                                              0.5  0.13                                                                              0.25 0.25                                                                              0.13                    S. aureus PC1 (Meth.sup.S)(lac+)                                                            ≦0.25                                                                       1   0.5  0.25                                                                              0.5 1    0.13                                                                              0.25 0.13                                                                              0.25                    S. aureus ATCC 13709 (Meth.sup.S)                                                           ≦0.25                                                                       1   0.5  0.25                                                                              0.25                                                                              0.5  0.06                                                                              0.13 0.06                                                                              0.13                    S. aureus Col (Meth.sup.R)(lac-)                                                            32   4   4    2   2   2    1   1    2   1                       S. aureus 76 (Meth.sup.R)(lac+)                                                             32   4   4    2   4   4    1   1    2   1                       S. aureus ATCC 33593 (Meth.sup.R)                                                           32   4   4    2   2   4    1   1    1   1                       S. aureus Spain #356 (Meth.sup.R)                                                           32   4   4    4   4   4    1   1    2   2                       S. haemolyticus 05 (Meth.sup.R)                                                             64   8   4    2   4   4    2   2    2   2                       E. faecalis ATCC 29212                                                                      ≦0.25                                                                       0.25                                                                              0.13 0.13                                                                              0.25                                                                              0.25 0.13                                                                              0.13 0.25                                                                              0.25                    E. faecium ATCC 35667                                                                       4    1   1    0.5 1   --   1   1    1   2                       E. faecium VanA (Van.sup.R)                                                                 4    2   2    1   2   2    1   2    2   4                       E. faecalis VanB (Van.sup.R)                                                                0.5  0.25                                                                              0.25 0.25                                                                              0.5 0.5  0.25                                                                              0.5  0.25                                                                              0.5                     E. faecium A491 (Amp.sup.R)                                                                 >128 16  16   16  16  16   32  32   >32 >32                     E. coli ATCC25992                                                                           ≦0.25                                                                       16  8    16  8   16   16  4    8   16                      P. aeruginosa ATCC 27853                                                                    1    >32 >32  >32 >32 >32  >32 32   >32 >32                     __________________________________________________________________________

Cmpd 1 (7R)-7-[(phenylacetyl)amino]-3-(3-isothioureidomethylpyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 2 (7R)-7-[(phenylacetyl)amino]-3-(4-isothioureidomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acidsalt

Cmpd 3 (7R)-7-[(Z)-2-(aminothiazol-4-yl) -2-(hydroxyimino)acetamido]-3-(3-isothioureidomethylpyrid-4-ylthio)-3-cephem -4-carboxylate,trifluoroacetic acid salt

Cmpd 4 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-isothioureidomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 5 (7R)-7-[(phenylacetyl)amino]-3-(4-isothioureidomethylthiazol-5-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 6 (7R)-7-[(phenylacetyl)amino]-3-[4-(3-pyrrolidinothiomethyl)-1,2,3-thiadiazol-5-ylthio]-3-cephem-4-carboxylate trifluoroaceticacid salt

Cmpd 7 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-isothioureidomethylthiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 8 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(cyclopentyloxyimino)acetamido]-3-(3-isothioureidomethylpyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 9 (7R)-7-[(phenylacetyl)amino]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 10 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 11 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[(methylaminoethylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 12 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[(guanidinoethylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 13 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(cyclopentyloxyimino)acetamido]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 14 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-guanidinoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 15 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-methylaminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 16 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 17 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(3-pyrrolidinothiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 18 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(N-methylglycyl)aminoethylthiomethylpyrid-4-ylthio]-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 19 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-1,1-dimethylethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 20 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiopyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 21(7R)-7-[(Z)-2-(2-amino-5-bromothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 22 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[(methylaminoethylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 23(7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 24 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(glycyl)aminoethylthiomethylpyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 25 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(N-formamidoyl)aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 26(7R)-7-[(Z)-2-phenyl-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 27 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(3-aminopropyl)thiomethylpyrid-4-ylthio]-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 28(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[(methylaminoethylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 29 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(glycyl)aminoethylthiomethylpyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 30 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiopyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 31 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(hiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(3-N-formamidoylaminopropylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 35 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(glycyl)aminopropylthiomethylpyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 36 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-aminoprop-1-ylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 37(7R)-7-[(phenylacetyl)amino]-3-(4-(2-aminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 38 (7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 39 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-carboxamidomethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylic acid

Cmpd 40 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-aminoethoxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 41 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-N-methylaminoethylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 42 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(pyrrolidin-3ylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 43 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(N-methylglycyl)aminoethylthiomethylpyrid-4-ylthio]-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 44 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(5-aminopentyloxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 45 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylsulfonylmethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 46 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(4-aminobutyloxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 47 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(3-aminopropyloxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 48 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(4-amino-2-butyn-1-yl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 49 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-aminopropyloxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 50 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(4-amino-2-Z-buten-1-yl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 51 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(3-aminopropylthio)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 52 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(piperidine-4-ylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 53 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylsulfinylmethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 54 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(N-acetamidinoyl)aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 55 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(5S-5-N,N-dimethylcarboxamidopyrrolidin-3-ylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 56 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2RS-2-amino-3-hydroxyprop-1-ylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 57 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(N-methylaminoethylaminoethylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 58 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2R-2-N,N-dimethylcarboxamido-2-aminoethylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 59 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2R-2-carboxamido-2-aminoethylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 60 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-isothioureidomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 61 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(N,N-dimethylcarboxamidomethylaminoethylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxyl ate,trifluoroacetic acid salt

Cmpd 62 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-amino-2-methylpropylthiomethy1)pyrid-4-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 63 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(N-formylaminoethylthiomethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 64 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(N-formylaminoethylthio)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 65 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2R-2-amino-3-hydroxyprop-1-ylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 66 (7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-[4-(2-aminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio]-3-cephem-4-carboxylate, trifluoroacetic acid salt

Cmpd 67 (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-aminoethylthiomethylpyrid-3-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 68 (7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-[4-(2-aminoethylthiomethyl)-1,3-thiazol-5-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt

Cmpd 69(7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-2-chloro-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

(Compound numbers correspond to those in MIC tables; data reported forchallenge with methicillin-susceptible S. aureus strain ATCC 13709.)

    ______________________________________                                        Compound: Survivors                                                           Dose      Vancomycin Imipenem 1     3    10                                   ______________________________________                                        10    mg/kg   10/10                                                           5     mg/kg   10/10                                                           2.5   mg/kg   4/10              10/10 9/10                                    1.25  mg/kg   2/10              8/10  7/10 10/10                              0.625 mg/kg   2/10       10/10  6/10  8/10 8/10                               0.3125                                                                              mg/kg              10/10  5/10  5/10 2/10                               0.156 mg/kg              10/10  2/10  0/10 2/10                               0.078 mg/kg              6/10              2/10                               0.039 mg/kg              3/10              1.29                               ED50 (mg/kg)                                                                            1.94       0.06     0.39  0.42                                      ______________________________________                                        Compound:   Survivors                                                         Dose        11       14      15    16    17                                   ______________________________________                                        40     mg/kg                                                                  20     mg/kg                                                                  10     mg/kg                                                                  5      mg/kg             10/10 5/10        6/10                               2.5    mg/kg    6/10     9/10  5/10  8/10  3/10                               1.25   mg/kg    5/10     3/10  2/10  5/9   5/10                               0.625  mg/kg    3/10     3/10  0/10  3/10  2/10                               0.3125 mg/kg    1/10     1/10  2/10  1/10  2/10                               0.156  mg/kg    1/10                 2/10                                     ED50 (mg/kg)                                                                              0.96     1.13    4.63  1.00  3.59                                 ______________________________________                                        Compound:   Survivors                                                         Dose        18       26      27    28    29                                   ______________________________________                                        5      mg/kg    6/10     9/10  7/10  7/10  7/10                               2.5    mg/kg    2/10     7/10  6/10  3/10  9/10                               1.25   mg/kg    2/10     5/10  2/10  4/10  9/10                               0.625  mg/kg    2/10     5/10  0/10  1/10  5/10                               0.3125 mg/kg    3/10     1/10  0/10  1/10  2/10                               ED50 (mg/kg)                                                                              4.44     1.06    2.64  3.01  1.26                                 ______________________________________                                        Compound:   Survivors                                                         Dose        30       32      33    39    56                                   ______________________________________                                        5      mg/kg    10/10    8/10  8/10  10/10 8/10                               2.5    mg/kg    10/10    5/10  9/10  6/10  8/10                               1.25   mg/kg    4/10     5/10  5/10  6/10  3/10                               0.625  mg/kg    2/10     1/10  4/10  3/10  3/10                               0.3125 mg/kg    0/10     0/10  0/10  1/10  2/10                               ED50 (mg/kg)                                                                              1.14     2.00    1.19  1.15  1.23                                 ______________________________________                                        Compound:   Survivors                                                         Dose        57       58      59    64    65                                   ______________________________________                                        5      mg/kg    9/10     5/10  10/10 8/10  8/10                               2.5    mg/kg    5/10     5/10  4/10  4/10  3/10                               1.25   mg/kg    4/10     2/10  4/10  4/10  1/10                               0.625  mg/kg    4/10     1/10  6/10  1/10  2/10                               0.3125 mg/kg    4/10     1/10  4/10  1/10  1/10                               ED50 (mg/kg)                                                                              1.08     4.10    0.81  2.27  3.10                                 ______________________________________                                        Compound:   Survivors                                                         Dose        66         67         68                                          ______________________________________                                        5      mg/kg    9/10       10/10    10/10                                     2.5    mg/kg    5/10       10/10    10/10                                     1.25   mg/kg    3/10       7/10     10/10                                     0.625  mg/kg    2/10       2/10     7/10                                      0.3125 mg/kg    3/10       2/10     4/10                                      ED50 (mg/kg)                                                                              1.65       0.79       0.39                                        ______________________________________                                    

EXAMPLES

The present invention will be more fully described in conjunction withthe following specific examples which are not to be construed in any wayas limiting the scope of the invention.

Example 1(7R)-7-[(phenylacetyl)amino]-3-(4-hydroxymethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

To a stirred solution of 1,3-dichloroacetone (6.85 g, 54 mmol) and ethyl3-mercaptopropionate (13.9 mL, 108 mmol) in anhydrous tetrahydrofuran(150 mL) was dropwise added triethylamine (15.0 mL, 108 mmol) at 0° C.and stirred at room temperature for 24 h. The solvent was evaporated andthe residue was partitioned between water and ethyl acetate. The ethylacetate phase was washed with 5% hydrochloric acid and then brine, driedover sodium sulfate, and concentrated to dryness, affording 17.4 g ofthe crude 1,3-bis(2-ethoxycarbonylethylthio)acetone.

A solution of 1,3-bis[(2-ethoxycarbonylethyl)thio]acetone (6.22 g, 19.3mmol), ethyl carbazate (2.40 g, 23.1 mmol) and a catalytic amount ofp-toluenesulfonic acid in anhydrous acetonitrile (50 mL) containingmolecular sieve was stirred for 2 days at room temperature. The reactionwas quenched with water/ethyl acetate and filtered. The filtrate wasextracted with ethyl acetate. The organic extract was washed withaqueous sodium bicarbonate, dried over sodium sulfate, and concentrated.The residue was triturated with hexane, affording 7.2 g of1,3-bis[(2-ethoxycarbonylethyl)thio]acetone ethoxycarbonylhydrazone. 1HNMR (CDCl₃) δ2.2-2.4 (m, 9H), 2.6-2.8 (m, 8H), 3.42 (s, 2H), 3.54 (s,2H), 4.1-4.3 (m, 6H), 8.82 (br s, 1H).

To a solution of the hydrazone (7.2 g, 17.6 mmol) in 1,2-dichloroethane(20 mL) was added thionyl chloride (3.9 mL, 53 mmol) at 0° C. andstirred overnight at room temperature. The mixture was concentrated to avolume of 10 mL, diluted with dichloromethane, washed with aqueoussodium bicarbonate, dried over sodium sulfate, and then concentrated todryness, affording 6.47 g of5-[(2-ethoxycarbonylethyl)thio]-4-[(2-ethoxycarbonylethyl)thiomethyl]-1,2,3-thiadiazole. 1H NMR (CDCl₃) δ1.2-1.3 (m, 6H), 2.60(t, 2H, J=7), 2.68 (t, 2H, J=7), 2.77 (t, 2H, J=7), 3.24 (t, 2H, J=7),4.07 (s, 2H), 4.1-4.2 (m, 4H)

To a solution of5-[(2-ethoxycarbonylethyl)thio]-4-[(2-ethoxycarbonylethyl)thiomethyl]-1,2,3-thiadiazole (1.0 g, 3.0 mmol) in dichloromethane (40mL) was added m-chloroperoxybenzoic acid in several portions at 0° C.until the starting material was disappeared. The mixture was thensequentially washed with saturated aqueous sodium thiosulfate, coldaqueous 1% sodium hydroxide and brine, and was concentrated to dryness.The oxidized product was treated with trifluoroacetic anhydride (2 mL)for 30 min at room temperature. After removal of trifluoroaceticanhydride under the reduced pressure, the mixture was stirred in ethylacetate/1% sodium hydroxide (30 ml/30 mL) for 30 min. The ethyl acetatelayer was concentrated to a volume of 5 mL and methanol (10 mL) wasadded. Excess sodium borohydride (200 mg) was added in portions at 0° C.while stirring. After 30 min, the reaction was quenched with dilutedhydrochloric acid. The mixture was extracted with ethyl acetate andconcentrated. The residue was purified by silica gel chromatography (1%methanol/dichloromethane) to provide 225 mg of5-[(2-ethoxycarbonylethyl)thio]-4-hydroxymethyl-1,2,3-th iadiazole. 1HNMR (CDCl₃) δ1.27 (t, 3H, J=8), 2.71 (t, 2H, J=7), 3.27 (t, 2H, J=7),4.16 (q, 2H, J=8), 5.03 (s, 2H).

To a solution of5-[(2-ethoxycarbonylethyl)thio]-4-hydroxymethyl-1,2,3-th iadiazole (225mg, 0.91 mmol) in anhydrous ethanol (20 mL) was added 0.5 M sodiummethoxide in methanol (1.6 mL). After 10 min, the mixture wasconcentrated, and was triturated with dichloromethane to obtain thesodium thiolate. A solution of the sodium thiolate and(7R)-7-[(phenylacetyl)amino]-3-trifluoromethanesulfonyloxy-3-cephem-4-carboxylate,4-methoxybenzyl ester (500 mg, 0.85 mmol) in tetrahydrofuran (10 mL) wasstirred at 0° C. for 30 min, and water (50 ml) was added. The reactionmixture was extracted with ethyl acetate and concentrated. The residuewas purified by silica gel chromatography (2% methanol/dichloromethane),affording 407 mg of the title compound. 1H NMR (CDCl₃) δ3.22 (d, 1H,J=18), 3.46 (d, 1H, J=18), 3.61 (d, 1H, J=16), 3.67 (d, 1H, J=16), 3.81(s, 3H), 4.95 (d, 1H, J=5), 5.02 (s, 2H), 5.23 (d, 1H, J=12), 5.28 (d,1H, J=12), 5.83 (dd, 1H, J=5, 8), 6.14 (d, 1H, J=8), 6.86 (d, 2H, J=9),7.2-7.4 (m, 7H).

Example 2(7R)-7-[(phenylacetyl)amino]-3-(4-chloromethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

To DMF (5 mL) was added thionyl chloride (85 mL, 1.71 mmol) and stirredfor 30 min at room temperature. A solution of(7R)-7-[(phenylacetyl)amino]-3-(4-hydroxymethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester (400 mg, 0.684 mmol) in DMF (1 mL) was added andstirred for an additional 30 min. The reaction was quenched with water,and the mixture was extracted with ethyl acetate. The organic extractwas washed with water and concentrated. The residue was purified bysilica gel chromatography (0.5% methanol/dichloromethane), affording 322mg of the title compound. 1H NMR (CDCl₃) δ3.20 (d, 1H, J=18), 3.47 (d,1H, J=18), 3.60 (d, 1H, J=16), 3.66 (d, 1H, J=16), 3.79 (s, 3H), 4.95(m, 3H), 5.22 (d, 1H, J=12), 5.25 (d, 1H, J=12), 5.85 (dd, 1H, J=5, 8),6.53 (d, 1H, J=8), 6.84 (d, 2H, J=9), 7.2-7.4 (m, 7H).

Example 3 (7R)-7-[(phenylacetyl)amino]-3-(4-isothioureidomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, trifluoroacetic acidsalt

A solution of(7R)-7-[(phenylacetyl)amino]-3-(4-chloromethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester (70 mg, 0.12 mmol), thiourea (7.6 mg, 0.1 mmol)and sodium iodide (15 mg, 0.1 mmol) in dry acetonitrile (3 mL) wasstirred at room temperature overnight. The mixture was concentrated andtriturated with dichloromethane to remove the unreacted startingmaterial. The residue was redissolved in acetone (2 mL) and filtered.The filtrate was concentrated to dryness, affording 60 mg of(7R)-7-[(phenylacetyl)amino]-3-(4-isothioureidomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, 4-methoxybenzyl ester,iodide salt as a yellowish solid. 1H NMR (acetone-d₆) δ3.56 (d, 1H,J=18), 3.66 (d, 1H, J=16), 3.72 (d, 1H, J=16), 3.80 (s, 3H), 3.86 (d,1H, J=18), 5.06 (d, 1H, J=15), 5.10 (d, 1H, J=15), 5.28 (m, 3H), 5.88(dd, 1H, J=5, 8), 6.91 (d, 2H, J=9), 7.2-7.4 (m, 7), 8.32 (d, 1H, J=8)

A mixture of (7R)-7-[(phenylacetyl)amino]-3-(4-isothioureidomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, 4-methoxybenzyl ester,iodide salt (60 mg), anisole (0.1 mL) and trifluoroacetic acid (1 mL)was stirred at 0 ° C. for 30 min, and then concentrated to dryness. Theresidue was sequentially triturated with dichloromethane (40 mL) andwater (0.5 mL) to afford 38 mg of the title compound. 1H NMR (DMSO-d₆)δ3.41 (d, 1H, J=18), 3.48 (d, 1H, J=14), 3.55 (d, 1H, J=14), 3.69 (d,1H, J=18), 4.86 (d, 1H, J=15), 4.94 (d, 1H, J=15), 5.14 (d, .1H, J=5),5.72 (dd, 1H, J=5, 8), 7.1-7.3 (m, 7H), 9.18 (d, 1H, J=8)

Example 4(7R)-7-amino-3-(4-chloromethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

To a solution of(7R)-7-[(phenylacetyl)amino]-3-(4-chloromethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester (604 mg, 1.68 mmol) and pyridine (0.271 mL, 3.36mmol) in dichloromethane (20 mL) was dropwise added a solution ofphosphorous pentachloride (208 mg, 2.68 mmol) in dichloromethane (10.7mL) and stirred at -10°C. for 2 h. After cooling to =40°C., isobutanol(1.55 mL) was added, and the resulting mixture was stirred at -10° C.for 4 h. The reaction was quenched with water, and the mixture wasextracted with dichloromethane. The organic extract was washed withsaturated aqueous sodium bicarbonate and concentrated. The residue waspurified by silica gel chromatography (10% methanol/dichloromethane),affording 599 mg of the title compound. ¹ H NMR (CD₃ OD) d 3.45 (d, 1H,J=18), 3.65 (d, 1H, J=18), 3.76 (s, 3H), 4.75 (1H, overlapped withwater), 4.98 (s, 2H), 5.05 (d, 1H, J=5), 5.23 (s, 2H), 6.82 (d, 2H,J=9), 7.26 (d, 2H, J=9).

Example 5 4-chloro-3-hydroxymethylpyridine

To a solution of 4-chloro-3-pyridyl carboxyaldehyde (140 mg, 1.0 mmol)in THF (1 mL) at 0° C. was added methanol (1 mL) followed by portionwiseaddition of sodium borohydride (75 mg, 2.0 mmol). After 1 hr, aceticacid (0.15 ml) was added and the reaction mixture was evaporated todryness with rotary evaporator at room temperature. The solid residuewas chromatographed on silica gel column (1% MeOH/dichloromethane) toafford 60 mg (42%) of the title compound. 1H NMR (CDCl₃) δ4.30 (br s,1H), 4.80 (s, 2H), 7.30 (d, 1H, J=5), 8.34 (d, 1H, J=5), 8.62 (s, 1H).

Example 6(7R)-7-[(phenylacetyl)amino]-3-(3-hydroxymethylpyrid-4-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

To a solution of 4-chloro-3-hydroxymethylpyridine (60 mg, 0.42 mmol) inDMF (1 mL) at room temperature under nitrogen was added potassiumthioacetate (71 mg, 0.63 mmol). After overnight stirring, the solventwas removed with rotary evaporator. The residue was washed with ethylether and taken up in 10% MeOH/dichloromethane. The insoluble materialwas filtered off and the filtrate was concentrated with rotaryevaporator. The residual material was dissolved in MeOH (3 mL) andaqueous sodium hydroxide was added (0.5 mL, 3 M). After an overnightreaction at room temperature the reaction was acidified with 1 Mhydrochloric acid, evaporated to dryness with rotary evaporator, andpartitioned between saturated sodium bicarbonate solution and ethylacetate. The organic layer was dried with sodium sulfate and the solventwas removed with rotary evaporator. The residue was dissolved in MeOHand(7R)-7-[(phenylacetyl)amino]-3-trifluoromethanesulfonyloxy-3-cephem-4-carboxylate,4-methoxybenzyl ester(240 mg, 0.42 mmol) was added followed by additionof dichloromethane. After an overnight reaction at room temperature thereaction was partitioned between 5% sodium bicarbonate solution andethyl acetate. Purification on silica gel (Chromatotron, 2%MeOH/dichloromethane) afforded the title compound (60 mg, 25%). 1H NMR(CDCl₃) δ3.15 (d, 1H, J=18), 3.55 (d, 1H, J=18), 3.63 (d, 1H, J=18),3.68 (d, 1H, J=18), 3.78 (s, 3H), 4.61 (d, 1H, J=13), 4.66(d, 1H, J=13),5.05 (d, 1H, J=5), 5.08 (d, 1H, J=13), 5.25 (d, 1H, J=13), 5.89 (dd, 1H,J=9, 5), 6.76 (d, 2H, J=8), 7.05 (m, 2H), 7.16 (d, 21H, J=8), 7.32 (m,5H), 8.40 (d, 1H, J=5), 8.48 (s, 1H).

Example 7(7R)-7-[(phenylacetyl)amino]-3-(3-chloromethylpyrid-4-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

To a solution of(7R)-7-[(phenylacetyl)amino]-3-(3-hydroxymethylpyrid-4-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester (112 mg, 0.194 mmol) and lithium chloride (14 mg,0.581 mmol) in DMF at 0° C. was added diisopropylethylamine (0.101 mL,0.581 mmol) and methanesulfonyl chloride (0.045 mL, 0.581 mmol). After45 min, the reaction mixture was partitioned between water and ethylacetate/hexane (v/v, 3/1). Purification on silica gel (Chromatotron, 2%MeOH/dichloromethane) afforded the title compound (52 mg, 45%). 1H NMR(CDCl₃) δ3.17 (d, 1H, J=18), 3.59 (d, 1H, J=18), 3.63 (d, 1H, J=18),3.68 (d, 1H, J=18), 3.77 (s, 3HH), 4.54 (d, 1H, J=13), 4.66 (d, 1H,J=13), 5.06 (d, 1H, J=5), 5.08 (d, 1H, J=13), 5.22 (d, 1H, J=13), 5.90(dd, 1H, J=9, 5), 6.76 (d, 2H, J=8), 6.95 (d, 1H, J=9), 7.07 (d, 1H,J=5), 7.15 (d, 2H, J=8), 7.30 (m, 5H), 8.41 (d, 1H, J=5), 8.45 (s, 1H).

Example 8(7R)-7-[(phenylacetyl)amino]-3-(3-isothioureidomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester, hydrochloride salt

To a solution of(7R)-7-[(phenylacetyl)amino]-3-(3-chloromethylpyrid-4-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester (52 mg, 0.087 mmol) in ethanol (0.4 mL) anddichloromethane (0.1 mL) was added thiourea (7 mg, 0.095 mmol) at roomtemperature. After overnight reaction the solvents were removed withrotary evaporator and the residue was triturated with ethyl ether. Thesolid precipitate was then dried in vacuum to afford the title compound(56 mg, 96%). 1H NMR (CDCl₃ /CD₃ OD) δ3.14 (d, 1H, J=10), 3.53 (d, 1H,J=10), 3.58 (s, 2H), 3.73 (s, 3H), 4.40 (s, 2H), 5.03 (d, 1H, J=5), 5.12(d, 1H, J=9), 5.15 (d, 1H, J=9), 5.75 (d, 1H, J=5), 6.74 (d, 2H, J=10),7.10 (d, 1H, J=5), 7.14 (d, 2H, J=10), 7.25 (m, 5H), 8.30 (d, 1H, J=5),8.45 (s, 1H).

Example 9(7R)-7-[(phenylacetyl)amino]-3-(3-isothioureidomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

To a solution of(7R)-7-[(phenylacetyl)amino]-3-(3-isothioureidomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester hydrochloride (56mg, 0.083 mmol) indichloromethane (1 mL) was added anisole (0.1 ml) followed by additionof trifluoroacetic acid (1 mL). After 30 min, the reaction mixture wasconcentrated with rotary evaporator and the residue was triturated withethyl ether. The precipitate was repeatedly washed by decantation withfresh portions of ethyl ether and dried in vacuum to afford the titlecompound (49 mg, 79%). 1H NMR (CD₃ OD) δ3.20 (d, 1H, J=18), 3.58 (d, 1H,J=13), 3.62 (d, 1H, J=13), 3.83 (d, 1H, J=18), 4.56 (d, 1H, J=10), 4.60(d, 1H, J=10), 5.25 (d, 1H, J=5), 5.78 (d, 1H, J=5), 7.28 (m, 5H), 7.43(d, 1H, J=5), 8.45 (d, 1H, J=s), 8.55 (s, 1H).

Example 10(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-chloromethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, 4-methoxybenzyl ester

To a suspension of(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetate sodium salt (2.11 g, 3.05 mmol) in DMF (4 mL) wasadded methanesulfonyl chloride (0.28 mL) at -60° C. and stirred at thesame temperature for 1.5 h. The solution was then added to a solution of(7R)-7-amino-3-(4-chloromethyl-1,2,3-thiadiazol-5-yl)thio-3-cephem-4-carboxylate,4-methoxybenzyl ester hydrochloride (920 mg, 1.88 mmol) anddiisopropylethylamine (0.4 mL) in DMF (2 mL) at -10° C. and stirred for1 h. The reaction mixture was poured into ice-water and the resultingprecipitate was collected by filtration. The filter cake was purified bysilica gel column chromatography (0.5% methanol/dichloromethane),affording 558 mg of the title compound. 1H NMR (CDCl₃) δ3.07 (d, 1H,J=18), 3.44 (d, 1H, J=18), 3.81(2H), 4.96 (2H), 5.06 (1H), 6.07 (1H),6.43 (s, 1H), 6.80 (br s, 1H), 6.88 (1H), 7.25-7.45 (40H).

Example 11(7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-isothioureidomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

A solution of(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-chloromethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester(221 mg, 0.19 mmol), sodium iodide (34 mg, 0.23mmol) and thiourea (14 mg, 0.18 mmmol) in acetonitrile (10 mL) wasstiired at 45° C. for 3 h. The resulting mixture was partitioned betweenwater and ethyl acetate. The ethyl acetate layer was washed with waterand concentrated. The residue was triturated to obtain theisothiouronium salt, which was subjected to the substantially the sameconditions for deprotection as used in Example 9, affording 27 mg of thetitle compound. 1H NMR (D₂ O) δ3.48 (d, 1H, J=18), 3.84 (d, 1H, J=18),5.39 (s, 1H), 5.90 (s, 1H), 7.16 (s, 1H).

Example 12 4-ethoxycarbonyl-5-[2-(phenylsulfonyl)ethylthio]thiazole

To a solution of potassium tert-butoxide (496 mg, 4.4 mmol) in 10 ml ofTHF was added a solution of ethyl isocyanoacetate (0.48 ml, 4.4 mmol) in5 ml of THF at -40° C. and the reaction mixture was continued to stirfor 10 min. After the reaction was cooled down to -60° C., a solutioncarbon disulfide in 5 ml of THF was added. The resulting mixture wasallowed to warm up to 0° C. and 2-iodoethyl phenyl sulfone (4.4 mmol)was added. The mixture was then stirred at refluxed condition for 5hours. After it was cooled down to room temperature, water and ethylacetate were added. The aqueous layer was adjusted to acidic conditionwith dilute hydrochloric acid and extracted with ethyl acetate. Thecombined organic layer was dried with anhydrous sodium sulfate and thesolvent was removed under reduced pressure. The crude residue waspurified by chromatography to give the title compound (890 mg, 56%). 1HNMR (CDCl₃) δ1.40 (t, 3H, J=7), 3.3-3.5 (m, 4H), 4.40 (q, 2H, J=7), 7.6(t, 2H, J=8), 7.70 (t, 1H, J=8), 7.93 (d, 2H, J=8), and 8.64 (S, 1H).

Example 13 4-hydroxymethyl-5-[2-(phenylsulfonyl)ethylthio]thiazole

To a solution of4-ethoxycarbonyl-5-[2-(phenylsulfonyl)ethylthio]thiazole (702 mg, 2mmol) in 20 ml of THF was added lithium borohydride (2M, 1 ml) andmethanol (0.16 ml, 4 mmol) at -30° C. The reaction mixture was allowedslowly warm up to room temperature and stirred overnight. The solventwas removed under reduced pressure and the crude residue was subjectedto chromatography by using dichloromethane and methanol as the eluent togive the title compound (400 mg, 65%). 1H NMR (CDCl₃) δ2.55 (s, 1H),3.00 (t, 2H, J=6), 3.40 (t, 2H, J=6), 4.71 (s, 2H), 7.6 (t, 2H, J=7),7.68 (t, 1H, J=7), 7.82 (d, 2H, J=7), and 8.80 (s, 1H).

Example 14(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-hydroxymethylthiazol-5-ylthio)-3-cephem-4-carboxylate,4 -methoxybenzyl ester

To a solution of 4-hydroxymethyl-5-[2-(phenylsulfonyl)ethylthio]thiazole(39 mg) in 1 ml of DMF was added potassium t-butoxide (14 mg) and theresulting mixture was continued to stir for 2 hours. After the reactionsolution was cooled down to -40° C., a solution of(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-trifluoromethanesulfonyloxy-3-cephem-4-carboxylate, 4-methoxybenzyl ester (104 mg) in 1.5 ml ofDMF was added. The mixture was allowed slowly warm up to 0° C., quenchedwith dilute aqueous ammonium chloride, and extracted with ethyl acetateand hexane. The solvent was removed under reduced pressure and the cruderesidue was purified by chromatography to give the title compound (61mg). 1H NMR (CDCl₃) δ3.28 (q, 2H, J=8), 3.82 (s, 3H), 4.72 (q, 2H, J=8Hz), 4.98 (d, 1H, J=4), 5.25 (q, 2H, J=8), 5.96 (q, 1H, J=4), 6.42 (s,1H), 6.85 (s, 1H), 6.93 (d, 2H, J=7), 7.20-7.42 (m, 33H), and 8.8 (s,1H).

Example 15(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-chloromethylthiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

To DMF (1 ml) was added thionyl chloride (0.016 ml) at 0° C. and theresulting mixture was continued to stir at the same temperature for 30min. The resulting solution was canulated to a solution of(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-hydroxymethylthiazol-5-ylthio)-3-cephem-4-carboxylate, 4-methoxybenzyl ester (61 mg) in 1 mlof DMF and continued to stir at the same temperature for 1 hour. Thereaction solution was diluted with ethyl acetate and hexane, and waswashed with water. The solvent was removed under reduced pressure andthe crude residue was purified by chromatography to give the titlecompound (43 mg). 1H NMR (CDCl₃) δ3.22 (q, 2H, J=12), 3.80 (s, 3H), 4.78(q, 2H, J=8), 5.04 (d, 1H, J=4), 5.30 (q, 2H, J=5), 6.00 (q, 1H, J=4),6.42 (s, 1H), 6.72 (s, 1H), 6.92 (d, 2H, J=7), 7.08 (d, 1H, J=4),7.20-7.45 (m, 32H), and 8.95 (s, 1H).

Example 16(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-isothioureidomethylthiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-chloromethylthiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester (43 mg) was dissolved in 2 ml of acetonitrile, towhich was added thiourea (4.5 mg) and sodium iodide (13 mg). Theresulting mixture was stirred at room temperature overnight. The solventwas removed under reduced pressure and the crude residue was purified byflash chromatography to give the title compound (40 mg). 1H NMR (CDCl₃/CD₃ OD) δ3.2 (q, 2H, J=12), 3.80 (s, 3H), 4.4 (q, 2H, J=12), 5.02 (d,1H, J=4 5.25 (q, 2H, J=5), 5.85 (d, 1H, J=4), 6.45 (s, 1H), 6.90 (d, 2H,J=7), 7.2-7.4 (m, 32H), and 9.00 (s, 1H).

Example 17(7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-isothioureidomethylthiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

(7R)-7-[(Z)-2-(N-triphenylmethylaminothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-isothioureidomethylthiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester(40 mg) was dissolved in 0.1 ml of anisole and 0.9ml of dichloroacetic acid. The resulting mixture was stirred at roomtemperature for 3 hours and then precipitated by addition of diethylether (100 ml). The precipitate was filtered and subjected to HP-20reverse phase chromatography to give the title compound. 1H NMR (D₂ O)δ3.60 (q, 2H, J=6), 4.60 (q, 2H, J=10), 5.20 (d, 1H, J=4), 5.80 (d, 1H,J=4), 6.90 (s, 1H), and 9.13 (s, 1H). IR (KBr) 997, 1042, 1180, 1349,1386, 1533, 1615, 1655, and 1768 cm⁻¹.

Example 18(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)aceticacid

To a solution of(Z)-2-(2-aminothiazol-4-yl)-2-(triphenylmethoxyimino)ace tic acid (5.81g, 13.47 mmol) in DMF (30 mL) at room temperature (1 mL) was addedN-chlorosuccinimide (1.80 g, 13.47 mmol). After overnight reaction thereaction mixture was poured into water (about 500 mL) and the resultingprecipitate was filtered, washed with water and then with ethyl acetateand dried in vacuum to afford 4.43 g (71%) of the title compound. 13CNMR (CDCl₃) δ108.5, 125.6, 126.2, 126.6, 12.3, 134.7 141.8, 146.5,162.1, 163.3.

Example 19(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylatediphenylmethyl ester

To a solution of 7-amino-3-chlorocephalosporanic acid diphenylmethylester toluenesulfonic acid salt (5.0 g, 8.72 mmol) in dry THF (100ml)was added pyridine (0.63 g. 10.0 mmol) at room temperature followedby addition of (Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetic acid (5.81 g, 13.47 mmol). The resulting slurry was cooledto -15° C. and additional pyridine (1.42 g, 22.5 mmol) was addedfollowed by dropwise addition of phosphorous oxychloride(1.64 g, 17.5mmol) while maintaining reaction temperature below -10° C. After 30 min.reaction ethyl acetate (200 mL) was added followed by addition of water(150 mL). Aqueous layer was thoroughly extracted with ethyl acetate andthe combined organic extracts were dried over sodium sulfate andconcentrated in vacuum to yield crude product which was purified byflash column chromatography on silica gel (ethyl acetate/hexane-3/1) toafford the title compound (5.37 g, 65%). 1H NMR (CDCl₃ /CD3OD) δ3.35 (d,1H, J=18), 3.68 (d, 1H, J=18), 5.07 (d, 1H, J=5), 5.80 (br s, 2H), 6.04(dd, 1H, J=9, 5), 7.03 (s, 1H), 7.06 (d, 1H, J=9), 7.22-7-50 (m, 25H).

Example 20(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-mercapto-3-cephem-4-carboxylatediphenylmethyl ester

To a solution of(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-chloro-3-cephem-4-carboxylate diphenylmethylester (4.0 g, 4.72 mmol) in DMF (30 mL) cooled to -20°C. was added inone portion powdered sodium hydrogen sulfide hydrate (1.1 g, 19.6 mmol).After 15 min the reaction mixture was poured into 0.5 M monosodiumphosphate (about 100 mL), extracted with ethyl acetate and the organiclayer was washed thoroughly with water. After concentrating in vacuumthe crude title product was obtained as yellow foam 3.8 g (95%). 1H NMR(CDCl₃ /CD3OD) δ3.38 (d, 1H, J=15) 4.43 (d, 1H, J=15), 5.03 (d, 1H,J=5), 5.80 (d, 1H, J=5), 5.99 (br s, 1H), 6.80 (s, 1H), 7.05-7.50 (m,25H).

Example 21 3-chloromethyl-4-chloropyridine hydrochloride

Thionyl chloride (0.714 mL, 9.78 mmol) was added at room temperature todry DMF (7 mL). After 30 min the above solution was cannulated into thesolution of 3-hydroxymethyl-4-chloropyridine (700 mg, 4.89 mmol) in DMF(3 mL). After 45 min, the product was precipitated by addition of dryether (100 ml), washed with ether, and dried in vacuum to yield 813 mg(84%) of the title compound. 1H NMR (CD₃ OD) δ5.00 (s, 2H), 8.31 (d, 1H,J=5), 8.99 (d, 1H, J=5), 9.18 (s, 1H).

Example 223-(N-tert-butoxycarbonylaminoethylthiomethyl)-4-chloropyridine

To a solution of 3-chloromethyl-4-chloropirydine hydrochloride (513 mg,2.59 mmol) in DMF (6 mL) at room temperature were added sodium iodide(386 mg, 2.59 mmol), diisopropylethylamine (1.12 mL, 6.47 mmol) and2-(N-tert-butoxycarbonylamino)ethanethiol (458 mg, 2.59 mmol). After 2h, the reaction mixture was partitioned between dilute HCl and ethylacetate. The organic layer was washed with water, dried over sodiumsulfate, and concentrated to yield 750 mg of the oily product (96%),which was used for the next step without further purification. 1H NMR(CDCl₃) δ1.43 (s, 9H), 2.61 (m, 2H), 3.35 (m, 2H), 3.81 (s, 2H), 4.90(br s, 1H), 7.35 (d, 1H, J=4), 8.40 (d, 1H, J=4), 8.57 (s, 1H).

Example 23(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-N-tert-butoxycarbonylaminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylatediphenylmethyl ester

To a solution of 7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-mercapto-3-cephem-4-carboxylatediphenylmethyl ester (650 mg, 0.777 mmol) in DMF (3 mL) was added3-(N-tert-butoxycarbonylaminoethylthiomethyl)-4-chloropy ridine (242 mg,0.80 mmol) at room temperature. After overnight reaction the reactionmixture was partitioned between water and ethyl acetate. The organiclayer was thoroughly washed with water, dried over sodium sulfate, andconcentrated to yield the crude product which was purified by radialchromatography on silica gel (dichloromethane/methanol; v/v, 50/1) toafford 220 mg of the title compound (26%). 1H NMR (CDCl₃ /CD₃ OD) δ1.23(s, 9H), 2.32 (t, 2H, J=6), 2.98 (d, 1H, J=18), 3.06 (m, 2H), 3.40 (d,1H, J=18), 3.46 (s, 2H), 5.03 (d, 1H, J=5), 5.52 (br s, 1H), 5.94 (d,1H, J=5), 6.80 (s, 1H), 6.90 (d, 1H, J=6), 7.00-7.22 (m, 25H), 8.01 (d,1H, J=6), 8.08 (s, 1H).

Example 24(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,methanesulfonic acid salt

To a solution of(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-N-tert-butoxycarbonylaminoethylthiomethylpyrid-4-ylthio)-3-cephem-4-carboxylate diphenylmethylester(1.0 g, 0.907 mmol) in dichloromethane (10 mL) and anisole (1.0 mL)was added at 0° C. trifluoroacetic acid (13 mL). After 1.5 hr thereaction mixture was concentrated in vacuum at room temperature and theoily residue was dissolved in 98% formic acid (20 mL). After 4 hr atroom temperature formic acid was removed in vacuum and the residue wasdissolved in water (25 mL). The insoluble material was removed bycentrifugation and the supernatant was purified on HP20 column byelution with water followed by 0.1 M ammonium acetate, water and finallyeluting the product with acetonitrile/water (1/4). The eluate wasconcentrated to about 1/10 original volume and the resulting precipitatewas filtered, washed with water and dried in vacuum to yieldzwitterionic product (260 mg). The methanosulfonate salt was prepared bysuspending the above material in water (15 mL) followed by addition ofmethanesulfonic acid (1.0 M in water, 0.98 eq) and acetonitrile (5 mL).After evaporation of the resulting solution to dryness the residue wasdissolved in water (30 mL), centrifuged to remove insoluble material andthe supernatant was lyophilized to produce the title compound (274 mg,44%). 1H NMR (D₂ O) δ3.11 (s, 3H), 3.19 (m, 2H), 3.52 (m, 2H), 3.67 (d,1H, J=17), 4.22 (d, 1H, J=17), 4.33 (s, 2H), 5.76 (d, 1H, J=4), 6.29 (d,1H, J=4), 7.93 (d, 1H, J=4), 8.78 (d, 1H, J=4), 8.87 (s, 1H).

Example 25 3-(N-tert-butoxycarbonylaminoethylthio)-4-chloropyridine

To a suspension of 4-chloropyridine hydrochloride (2 g) in 40 mL of dryTHF was added a freshly prepared LDA (2.5 eq.) at -70° C. and theresulting mixture was continued to stir at the same temperature for 4hours. A solution of N, N'-di(tert-butoxycarbonyl)cystamine (2.5 g, 0.5eq.) in 10 ml of THF was cannulated to the above solution. The reactionmixture was allowed to warm up to 0° C., quenched with water and thenextracted with ethyl acetate. The solvent was removed under reducedpressure and the crude residue was purified by flash chromatography togive of the title compound(0.95 g, 50%) as a white solid. 1H NMR (CDCl₃)δ1.42 (s, 9H), 3.15 (t, 2H, J=7), 3.38 (t, 2H, J=7), 4.95 (s, 1H), 7.32(d, 1H, J=6), 8.35 (d, 1H, J=6), and 8.60 (s, 1H).

Example 26(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-[3-(N-tert-butoxycarbonylaminoethylthio)pyridyl-4-thio]-3-cephem-4-carboxylate,diphenylmethyl ester

To a stirring solution of(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-mercapto-3-cephem-4-carboxylate,diphenylmethyl ester (6.7 g, 7.8 mmol) in 20 ml of dry DMF was added3-(N-tert-butoxycarbonylaminoethylthio)-4-chloropyridine (2.3 g, 7.8mmol) at -20° C. The reaction mixture was allowed to slowly warm up toroom temperature and stirred overnight. The reaction mixture was dilutedwith ethyl acetate and hexane and washed with water. The solvent wasremoved under reduced pressure and the crude residue was purified bychromatography to give the title compound (6 g, 68%) as a yellow solid.1H NMR (CDCl₃) δ1.40 (s, 9H), 2.75 (t, 2H, J=7), 3.10 (t, 2H, J=7), 3.15(d, 1H, J=14), 3.60 (d, 1H, J=14), 4.95 (s, 1H), 5.20 (d, 1H, J=4), 5.90(s, 2H), 6.25 (q, 1H, J=4), 6.85 (d, 1H, J=4), 6.90 (s, 1H), 7.15-7.4(m, 26H), 8.1 (s, 1H), 8.21 (d, 1H, J=7).

Example 27(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylthiopyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

To a solution of(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-[3-(N-tert-butoxycarbonylaminoethylthio)pyridyl-4-thio]-3-cephem-4-carboxylate, diphenylmethyl ester(6 g) in 10 ml of dichloromethane and 1 ml of anisole was added 10 ml oftrifluoroacetic acid at 0° C. and the resulting solution was stirred atroom temperature for 1 hour. After the solvent was removed, the residuewas redissolved in 20 ml formic acid and stirred at room temperatureovernight. The solvent was removed under reduced pressure. The residuewas dried over high vacuum and then triturated with ethyl acetate. Theresulting solid was subjected to reverse phase chromatography onAmberchrom column (0.1% aqueous trifluoroacetic acid/acetonitrile) togive the title compound(1.5 g, 48%). 1H NMR (D₂ O) δ3.5-3.7 (m, 4H), 3.8(d, 1H, J=14), 4.4 (d, 1H, J=14), 5.84 (d, 1, J=4), 6.4 (d, 1H, J=4),7.65 (d, 1H, J=6), 8.82 (d, 1H, J=6), and 9.02 (s, 1H). IR (KBr) 778,1042, 1173, 1541, 1610, 1780, 3187 cm⁻¹.

Example 28(7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(triphenylmethoxyoxyimino)acetamido]-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester

To a solution of4-[(2-tert-butoxycarbonylaminoethyl)thiomethyl]-5-(2-ethoxycarbonylethyl)thio-1,2,3--1,2,3-thiadiazole(181 mg, 0.44 mmol) in ethanol (5 mL) was added sodium methoxide inmethanol (0.55M, 1 mL), and concentrated. The residue was trituratedwith hexane-ethyl acetate(9:1), mixed with(7R)-7-[[(Z)-2-(2-aminothiazol-4-yl)-2-(triphenylmethoxyimino)acetyl]amino]-3-chloro-3-cephem-4-carboxylate, diphenylmethylester (200 mg, 0.25 mmol), and dissolved in a mixed solvent of ethanoland dichloromethane. The solution was stirred at room temperature for 16h, and concentrated. The crude was purified by silica gel columnchromatography (3% methanol/dichloromethane), affording 100 mg of thetitle compound. 1H NMR (CDCl₃) δ1.46 (9H), 2.68 (2H), 3.19 (d, 1H,J=18), 3.33 (2H+1H), 4.03 (2H), 4.88 (br s, 1H), 5.09 (1H), 5.93 (br s,2H), 6.44 (s, 1H), 7.03 (1H), 7.20-7.45 (25H).

Example 29(7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,dichloroacetic acid salt

(7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(triphenylmethoxyoxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,diphenylmethyl ester (82 mg, 0.076 mmol) was dissolved in 0.8 mL ofdichloroacetic acid containing 5% anisole and stirred at roomtemperature overnight. Excess diethyl ether was added and the resultingprecipitate was collected by filtration. The filter cake was purified byHP-20 reverse phase column chromatography, affording 16 mg of the titlecompound. 1H NMR (DMSO-d₆) δ2.64 (2H), 2.90 (2H), 3.4 (1H, overlappedwith water), 3.82 (d, 1H, J=17), 4.04 (d, 1H, J=15), 4.14 (d, 1H, J=15),5.17 (d, 1H, J=5 ), 5.74 (dd, 1H, J=5, 8), 6.64 (s, 1H), 7.08 (s, 1H),9.50 (d, 1H, J=8).

Example 30 3-(2-N-tert-butoxycarbonylaminoethoxymethyl)-4-chloropyridine

A biphasic mixture of 4-chloro-3-chloromethylpyridine hydrochloride (396mg, 2 mmol), N-tert-butoxycarbonylaminoethanol (132 mg, 2 mmol) andbenzyl triethyl ammonium bromide (544 mg, 2 mmol) in toluene (20 mL) and50% sodium hydroxide aqueous solution was vigorously stirred at roomtemperature for 24 h. The organic layer was taken and concentrate toafford 490 mg of the title compound. 1H NMR (CDCl₃) δ1.46 (s, 9H), 3.39(2H), 3.65 (2H), 4.64 (s, 2H), 4.90 (br s, 1H), 7.33 (d, 1H, J=5), 8.46(d, 1H, J=5), 8.64 (s, 1H).

Example 31(7R)-7-[(Z)-2-(2-amino-5-chlorothiazo-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-(2-tert-butoxycarbonylaminoethoxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,diphenylmethyl ester

Under substantially the same conditions as used in Example 26,(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-mercapto-3-cephem-4-carboxylate,diphenylmethyl ester was reacted with3-(2-N-tert-butoxycarbonylaminoethoxymethyl)-4-chloropyridine to affordthe title compound. 1H NMR (CDCl₃) δ1.42 (9H), 3.11 (d, 1H, J=18), 3.24(2H), 3.40 (2H), 3.46 (d, 1H, J=18), 4.30 (2H), 4.83 (br s, 1H), 5.17(1H), 5.72 (br s, 2H), 6.20 (1H), 6.99 (s, 1H), 7.25-7.45 (25H), 8.28(1H), 8.33 (1H).

Example 32(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-aminoethoxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-(2-tert-butoxycarbonylaminoethoxymethyl)pyrid-4-ylthio)-3-cephem-4-carboxylate, diphenylmethylester was subjected to deprotection under substantially the sameconditions as used in Example 27, affording the title compound. 1H NMR(D₂ O) δ3.35 (2H), 3.45 (d, 1H, J=18), 3.91 (2H), 4.00 (d, 1H, J=18),5.52 (d, 1H, J=5), 6.04 (d, 1H, J=5), 7.73 (d, 1H, J=6), 8.57 (d, 1H,J=6), 8.65 (s, 1H).

Example 333-(N-tert-butoxycarbonylaminoethylsulfonylmethyl)-4-chloropyridine

To a solution of3-(N-tert-butoxycarbonylaminoethyl)thiomethyl)-4-chloropyridine (302 mg,1 mmol) in a mixed solvents of ethyl acetate (10 mL) and methanol (5 mL)was added methanesulfonic acid (144 mg, 1.5 mmol). 3-chloroperoxybenzoicacid (700 mg) was added and stirred at room temperature for 5 h. Thereaction was quenched with saturated sodium thiosulfate aqueoussolution. The solution was neutralized with 10% sodium hydroxide andextracted with ethyl acetate. The ethyl acetate extract was dried oversodium sulfate and concentrated to dryness, affording 286 mg of thetitle compound. 1H NMR (CDCl₃) δ1.44 (9H), 3.22 (2H), 3.66 (2H), 4.50(s, 2H), 5.21 (br s,1H), 7.42 (d, 1H, J=5), 8.54 (s,1H, J=5), 8.72 (1H).

Example 34(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-N-tert-butoxycarbonylaminoethylsulfonylmethylpyrid-4-ylthio)-3-cephem-4-carboxylate,diphenylmethyl ester

Under substantially the same conditions as used in Example 26,(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-mercapto-3-cephem-4-carboxylate,diphenylmethyl ester was reacted with3-(N-tert-butoxycarbonylaminoethylsulfonylmethyl)-4-chloropyridine toafford the title compound. 1H NMR (CDCl₃) δ1.44 (9H), 3.20 (3H), 3.66(3H), 4.48 (s, 2H), 5.19 (1H), 7.25-7.45 (25H), 8.50 (s,1H, J=5), 8.70(1H).

Example 35(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylsulfonylmethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-N-tert-butoxycarbonylaminoethylsulfonylomethylpyrid-4-ylthio)-3-cephem-4-carboxylate,diphenylmethyl ester was subjected to deprotection under substantiallythe same conditions as used in Example 27, affording the title compound.1H NMR (D₂ O) δ3.47 (d, 1H, J=18), 3.3.70 (2H), 3.85 (2H), 4.02 (d, 1H,J=18), 5.07(s, 2H), 5.52 (d, 1H, J=5), 6.03 (d, 1H, J=5), 7.79 (d, 1H,J=6), 8.64 (d, 1H, J=6), 8.76 (s, 1H).

Example 36 3-(4-N-tert-butoxycarbonylaminobutyn-1-yl)-4-chloropyridine

4-chloro-3-chloromethylpyridine hydrochloride was reacted with3-t-BOC-amino-1-propyne under the phase transfer condition described inExample 30, affording the title compound. 1H NMR (CDCl₃) δ1.4-1.6 (9H,rotomeric mixture), 4.0-4.2 (2H), 7.32 (d, 1H, J=5), 8.43 (d, 1H, J=5),8.53 (s, 1H).

Example 37(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-(4-N-tert-butoxycarbonylaminobutyn-1-yl)pyrid-4-ylthio)-3-cephem-4-carboxylate,diphenylmethyl ester

Under substantially the same conditions as used in Example 26,(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-mercapto-3-cephem-4-carboxylate,diphenylmethyl ester was reacted with 3-(4-N-tert-butoxycarbonylaminobutyn-1-yl)-4-chloropyridine to afford the titlecompound. 1H NMR (CDCl₃) δ1.4-1.8 (9H), 3.10 (d, 1H, J=18), 3.47 (d, 1H,J=18), 3.85 (2H), 5.16 (1H), 5.62 (br s, 2H), 6.13 (1H), 7.00 (1H),7.2-7.5 (25H), 8.35 (2H).

Example 38(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(4-amino-2-butyn-1-yl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-(4-N-tert-butoxycarbonylaminobutyn-1-yl)pyrid-4-ylthio)-3-cephem-4-carboxylate, diphenylmethylester was subjected to deprotection under substantially the sameconditions as used in Example 27, affording the title compound. 1H NMR(D₂ O) δ3.46 (d, 1H, J=18), 4.02 (d, 1H, J=18), 4.16 (2H), 4.70 (2H),5.52 (d, 1H, J=5), 6.03 (d, 1H, J=5), 7.76 (d, 1H, J=6), 8.66 (d, 1H,J=6), 8.78 (s, 1H).

Example 39(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-(4-amino-2-(Z)-buten-1-yl)pyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-(4-tert-butoxycarbonylamino-2-(Z)-buten-1-yl)pyrid-4-ylthio)-3-cephem-4-carboxylate,diphenylmethyl ester was subjected to deprotection under substantiallythe same conditions as used in Example 27, affording the title compound,as a mixture with the 1-butenyl isomer. 1H NMR (D₂ O) δ3.46 (d, 1H,J=18), 3.91 (2H), 4.02 (d, 1H, J=18), 4.56 (2H), 5.52 (d, 1H, J=5), 5.63(2H), 6.03 (d, 1H, J=5), 7.76 (d, 1H, J=6), 8.65 (d, 1H, J=6), 8.76 (s,1H).

Example 403-(N-tert-butoxycarbonylaminoethylsulfenylmethyl)-4-chloropyridine

To a solution of3-(N-tert-butoxycarbonylaminoethyl)thiomethyl)-4-chloropyridine (687mg,2.26 mmol) in methylene chloride (10 mL) at 0° C. was added3-chloroperoxybenzoic acid (467, 2.72 mmol). After overnight reaction atroom temperature reaction mixture was partitioned betweendichloromethane and diluted sodium bicarbonate solution. The organiclayer was dried over sodium sulfate and concentrated. Crystallization ofthe crude from ethyl acetate/hexane afforded the title compound. 1H NMR(CDCl₃) δ1.40 (s, 9H), 2.90 (m, 1H), 3.01 (m, 1H), 3.59 (m, 2), 4.00 (d,1H, J=13), 4.22 (d, 1H, J=13), 5.30 (br. s, 1H), 7.40 (d, 1H, J=4), 8.45(d, 1H, J=4), 8.57 (s, 1H)

Example 41(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(3-aminoethylsulfenylmethylpyrid-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

Under substantially the same conditions as used in Example 43,(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-mercapto-3-cephem-4-carboxylate,diphenylmethyl ester was reacted with3-(N-tert-butoxycarbonylaminoethylsulfenylmethyl)-4-chloropyridine toafford (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-(3-N-tert-butoxycarbonylaminoethylsulfenylmethylpyrid-4-ylthio)-3-cephem-4-carboxylate,diphenylmethyl ester, which was then subjected to deprotection undersubstantially the same conditions as used in Example 35, affording thetitle compound. 1H NMR (D₂ O) δ2.90 (s, 3H), 3.20-3.35 (m, 1H),3.50-3.70 (m, 4H), 4.00 (dd, 1H, J=18, 3), 4.55 (dd, 1H, J=13, 3), 4.65(dd, 1H, J=13, 3), 5.51 (m, 1H), 6.02 (m, 1H), 7.75 (m, 1H), 8.60 (m,2H).

Example 42 (7R)-7-{[2-[N,N'-bis-(t-butoxycarbonyl)guadininolethylthio]acetyllamino)-3-(4-chloromethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester

To a solution of (7R)-7-amino-3-(4-chloromethyl-1,2,3-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, 4-methoxybenzylester (485 mg, 0.794 mmol) and {2-[N^(W),N^(W') -bis- (t-butoxycarbonyl)guadinino]ethyl} thioacetic acid (329 mg, 0.873 mmol) were addedphosphorous oxychloride (0.103 mL, 1.11 mmol) and diisopropylethyl amine(0.55 mL, 3.18 mmol), and was stirred at -10° C. for 16 h. Water wasadded, and the mixture was extracted with ethyl acetate. The organicextract was washed with aqueous sodium bicarbonate and thenconcentrated. The residue was purified by silica gel chromatography (1%methanol/dichloromethane), affording 440 mg of the title compound. 1HNMR (CD₃ OD) δ1.44 (s, 9), 1.47 (s, 9), 2.79 (m, 2), 3.30 (2, overlappedwith solvent), 3.35 (d, 1, J=18), 3.57 (m, 2), 3.64 (d, 1, J=18), 3.77(s, 3), 4.96 (s, 2), 5.15 (d, 1, J=5), 5.20 (d, 1, J=12), 5.23 (d, 1,J=12), 5.77 (d, 1, J=5) 6.81 (d, 2, J=9), 7.25 (d, 1, J=8).

Example 43(7R)-7-{[2-[N,N'-bis-(t-butoxycarbonyl)guadinino]ethylthio]acetyl]amino}-3-(4-isothiouroniummethylthiomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester, iodide salt

A solution of(7R)-7-[2-[N,N'-bis-(t-butoxycarbonyl)guadinino]ethylthio]acetyl]amino}-3-(4-chloromethyl-1,2,3--1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester (200 mg, 0.24 mmol), thiourea (18 mg, 0.24 mmol)and sodium iodide (35 mg, 0.24 mmol) in acetonitrile (3 mL) was stirredat room temperature overnight. The mixture was concetrated andtriturated with dichloromethane. The residue was redissolved in acetone(2 mL) and filtered. The filtrate was concentrated to dryness, affording200 mg of the title compound. ¹ H NMR (CDCl₃) δ1.45 (s, 18), 2.81 (m,2), 3.15 (d, 1, J=18) 3.36 (d, 1, J=15), 3.55 (d, 1, J=15), 3.62 (m, 2),3.80 (s, 3), 3.83 (d, 1, J=18), 4.80 (d, 1, J=15), 4.86 (d, 1, J=15),5.08 (d, 1, J=5), 5.23 (d, 1, J=12), 5.27 (d, 1, J=12), 5.58 (dd, 1,J=5, 8) 6.85 (d, 2, J=9), 7.37 (d, 2, J=9), 8.24 (d, 1, J=8).

Example 44(7R)-7-{[(2-guadininoethylthio)acetyl]amino}-3-(4-isothiouroniummethylthiomethyl-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylic acid, 2,2-dichloroacetate salt

A solution of(7R)-7-{[2-[N,N'-bis-(t-butoxycarbonyl)guadinino]ethylthio]acetyl]amino}-3-(4-isothiouroniummethylthiomethyl-1,2,3--1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,4-methoxybenzyl ester, iodide salt (200 mg) and anisole (0.1 mL) indichloroacetic acid (2 mL) was stirred at room temperature for 16 h. Themixture was precipitated with diethylether/hexane, filtered, andvacuum-dried to afford 70 mg of the title compound.

Using substantially the same methods as described in the Examples above,the following additional compounds were prepared:

Example 45(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 46(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 47(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 48(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminopropylthio)pyrid-4-ylthio]3-cephem-4-carboxylate,trifluoroacetic acid salt Example 49(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 50(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 51(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 52(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 53(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 54(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 55(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 56(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 57(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 58(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 59(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 60(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 61(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 62(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 63(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 64(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 65(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 66(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 67(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 68(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 69(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 70(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 71(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 72(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 73(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 74(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 75(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 76(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 77(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboximidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 78(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetimido]-3-[3-(2-amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 79(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetimido]-3-[3-(2-guanidino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 80(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetimido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 81(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 82(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 83(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 84(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 85(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidinopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 86(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 87(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 88(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 89(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 90(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 91(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-guanidino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 92(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 93(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 94(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 95(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 96(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 97(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 98(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 99(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 100(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 101(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 102(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 103(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 104(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 105(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 106(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 107(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 108(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 109(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 110(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 111(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 112(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 113(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 114(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 115(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 116(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 117(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-25aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate, trifluoroaceticacid salt Example 118(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 119(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 120(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 121(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 122(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 123(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 124(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 125(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboxamidoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 126(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 127(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 128(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthio)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 129(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 130(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 131(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 132(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 133(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidinopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 134(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)aminopropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 135 (7R)-7-[(Z)-2-(²-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 136(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 137(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 138(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 139(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-guanidino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 140(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-N-(iminomethyl)amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 141(7R)-7-amino-3-[4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio]-3-cephem-4-carboxylate,benzhydryl ester, p-toluenesulfonic acid salt

To a solution of4-[(2-tert-butoxycarbonylaminoethyl)thiomethyl]-5-(2-ethoxycarbonylethyl)thio-1,2,3-thiadiazole(650 mg, 1.65 mmol) in isopropyl alcohol (5 mL) was added sodiummethoxide in methanol (0.55M, 1 mL), and the mixture was concentrated.The residue was mixed with(7R)-7-amino-3-methanesufonyloxy-3-cephem-4-carboxylate, diphenylmethylester, p-toluenesulfonic acid salt (870 mg, 1.37 mmol) and then treatedwith a mixed solvent of ethyl acetate (25 mL) and water (10 mL)containing sodium bicarbonate (350 mg, 4.11 mmol). The biphasic reactionmixture was saturated with sodium chloride and stirred at roomtemperature for 1 h. The organic layer was removed and concentratedafter addition of p-toluenesulfonic acid (260 mg, 1.37 mmol). The crudewas triturated with diethyl ether to afford 1.10 g of the titlecompound. 1H NMR (CDCl3) δ1.42 (9H), 2.28 (3H), 2.60 (4H), 3.28 (1H),3.40 (br d, 1H), 3.94 (2H), 4.87 (2H), 5.02 (1H), 6.96-7.65 (15H).

Example 142 Methyl 6-tert-butoxycarbonylamino-2-pyridinecarboxylate

To a suspension of 2,6-pyridinedicarboxylic acid (8.35 g, 50 mmol) andtriethylamine (5.05 g, 50 mmol) in tert-butyl alcohol (ca. 60 mL) wasdropwise added diphenylphosphoryl azide (13.8 g, 50 mmol), and themixture was stirred at room temperature until the reaction mixturebecame a clear solution. The solution was then slowly heated andrefluxed for 2.5 hour. After cooling, the mixture was partitionedbetween ethyl acetate/hexane (1:1, v/v) and 1% aqueous sodium hydroxide.The organic layer was washed again with 1% aqueous sodium hydroxide. Allthe aqueous solutions were combined and neutralized to pH=5 withconcentrated hydrochloric acid in the presence of ethyl acetate. Theethyl acetate layer was washed with brine, dried over sodium sulfate andconcentrated to obtain 5.0 g of6-tert-butoxycarbonylamino-2-pyridinecarboxylic acid.

This acid (1.38 g, 5.8 mmol) was added to an ethyl acetate solution (20mL) of Vilsmeier reagent (prepared from phosphorus oxychloride (1.06 g,6.9 mmol) and N,N-dimethylformamide (0.51 g, 6.9 mmol)) and stirred at0° C. for 1 h. Anhydrous methanol (10 mL) was added, and the mixture wasstirred for an additional 15 min. The mixture was then concentrated andpartitioned between ethyl acetate and saturated aqueous sodiumbicarbonate. The organic phase was washed with 1% aqueous lithiumchloride and then brine, dried over sodium sulfate, and concentrated toafford 1.4 g of the title compound as a white crystalline solid. 1H NMR(CDCl3) δ1.51 (9H), 3.98 (3H), 7.54 (br s, 1H), 7.80 (2H), 8.16 (1H).

Example 143 Methyl 6-tert-butoxycarbonylamino-2-pyridineglyoxylate

To a solution of methyl 6-tert-butoxycarbonylamino-2-pyridinecarboxylate(5.3 g, 21 mmol) and methyl methylsufinylmethyl sulfide (3.7 g, 29 mmol)in N,N-dimethylformamide (15 mL) was added sodium hydride (60% in oil,1.3 g, 34 mmol) at 0° C., and the mixture was stirred at roomtemperature for 2 h. Diethyl ether (100 mL) was added and the mixturewas stirred for 30 min. The resulting precipitate was collected,dissolved in water, neutralized to pH=5 with 6% hydrochloric acid, andextracted with diethyl ether. The ether extract was dried andconcentrated to afford 3.5 g of the adduct.

This adduct was dissolved in dichloromethane (10 mL), and pyridine (3.1g, 40 mmol) and trifluoroacetic anhydride (4.2 g, 20 mmol) were added at0° C. After stirring at room temperature for 30 min, 0.5 M sodiummethoxide in methanol (15 mL) was added. The mixture was then stirredfor 1 h and concentrated. Extraction with ethyl acetate from aqueoussodium bicarbonate followed by evaporation afforded 3.7 g of the crudedithioorthoester.

The above crude (10 g) was dissolved in acetic acid (30 mL) and sodiumperborate monohydrate (8.4 g, 84 mmol) was added. The resultingsuspension was stirred for 2.5 h. The solvent was evaporated and ethylacetate was added. The resulting solution was then washed with aqueoussodium bicarbonate, concentrated, and chromatographed (silica gel,dichloromethane) to afford 3.2 g of the title compound. 1H NMR (CDCl3)δ1.53 (9H), 3.98 (3H), 7.32 (br s, 1H), 7.77 (d, 1H), 7.86 (t, 1H), 8.24(d, 1H)

Example 1442-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)aceticacid

To a solution of methyl 6-tert-butoxycarbonylamino-2-pyridineglyoxylate(1.6 g, 5.7 mmol) in 95% ethanol were added hydroxylamine hydrochloride(0.6 g, 8.6 mmol) and pyridine (0.96 mL, 9.7 mmol), and the mixture wasstirred at room temperature for 1.5 h. The solvent was removed in vacuoand the oily residue was partitioned between ethyl acetate and water.The organic solution was washed with 3% hydrochloric acid and thenwater, dried over sodium sulfate, and concentrated to give 1.63 g of thecrude hydroxyimino ester.

The crude ester was dissolved in dichloromethane and cooled to 0° C.Trityl chloride (1.87 g, 6.7 mmol) and triethylamine (0.84 mL, 6.7 mmol)were added and stirred for 2 h. The mixture was diluted withdichloromethane, washed with 1% hydrochloric acid and concentrated toafford 3.1 g of the tritylated ester. Crystallization from hexane-ethylacetate provided the syn-oxyimino product that was free from theanti-isomer.

The above ester (4.25 g, 7.91 mmol) was dissolved in a mixed solvent ofisopropyl alcohol, tetrahydrofuran and water (10:5:1, v/v/v) containingsodium hydroxide (0.64 g, 16 mmol) and was stirred at 65° C. for 1 h.The reaction mixture was concentrated to dryness, triturated withhexane, and partitioned between dichloromethane and diluted hydrochloricacid. The organic layer was concentrated, affording 3.7 g of the titlecompound. 1H NMR (CDCl3) δ1.53 (9H), 7.05-7.50 (16H), 7.55 (1H), 7.64(1H).

Example 145(7R)-7-[(Z)-2-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester

To a solution of(7R)-7-amino-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester, p-toluenesulfonic acid salt (700 mg, 0.81 mmol) and2-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)aceticacid (750 mg, 1.46 mmol) in anhydrous tetrahydrofuran (30 mL) weresequentially added phosphorous oxychloride (0.10 mL, 1.22 mmol) anddiisopropylethylamine (0.38 mL, 3.24 mmol) at -40° C. The resultingsolution was stirred for 30 min and the reaction was quenched with 0.1%hydrochloric acid. The mixture was extracted with ethyl acetate andconcentrated. The residue was then immediately chromatographed on silicagel (0.5% methanol/dichloromethane), affording 445 mg of the titlecompound. 1H NMR (CDCl3) δ1.42 (9H), 1.50 (9H), 2.68 (2H), 3.20 (d, 1H,J=18), 3.34 (2H), 3.45 (d, 1H, J=18), 4.04 (t, 2H), 4.88 (br s, 1H),5.18 (d, 1H, J=5), 6.21 (1H), 7.04 (s, 1H), 7.15 (d, 1H, J=8), 7.21-7.47(25H), 7.53 (t, 1H, J=8), 7.81 (d, 1H, J=8).

Example 146(7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

A mixture of(7R)-7-[(Z)-2-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,2,3-thiadiazol-5-ylthio)-3-cephem-4-carboxylate, benzhydryl ester (125 mg, 0.106 mmol) and phenol (625 mg) indichloroacetic acid (0.5 mL) was stirred at 48° C. for 1 h. Aftercooling, diethyl ether was added. The resulting precipitate wascollected, dissolved in water, and filtered. The filtrate was subjectedto reverse phase chromatography on Amberchrom column (from 0 to 50%water (containing 0.1% trifluoroacetic acid)/acetonitrile) to give 68 mgof the title compound. 1H NMR (D₂ O) δ2.91 (2H), 3.30 (2H), 3.49 (d, 1H,J=18), 3.75 (d, 1H, J=18), 4.27 (2H), 5.38 (1H), 5.90 (1H), 6.98 (d, 1H,J=8), 7.11 (d, 1H, J=8), 7.93 (t, 1H, J=8).

Example 147 4-Chloromethyl-5-(2-phenylsulfonylethyl)thio-1,3-thiazole

Thionyl chloride (1.5 mL) was mixed with N,N-dimethylformamide (15 mL)and stirred at room temperature for 30 min. The resulting solution wasthen transferred via syringe to a solution of4-hydroxymethyl-5-(2-phenylsulfonylethyl)thio-1,3-thiazole (1.69 g, 5.36mmol) in N,N-dimethylformamide (5 mL). After 2 h, the mixture waspartitioned between ethyl acetate and water. The organic solution waswashed with 5% aqueous sodium bicarbonate, dried over sodium sulfate,and concentrated to dryness, affording 1.78 g of the title compound. 1HNMR (CDCl3) δ3.05 (2H), 3.35 (2H), 4.70 (2H), 7.50-7.85 (5H), 8.82 (s,1H).

Example 1484-[(2-tert-butoxycarbonylaminoethyl)thiomethyl]-5-(2-phenylsulfonylethyl)thio-1,3-thiazole

A mixture of 4-chloromethyl-5-(2-phenylsulfonylethyl)thio-1,3-thiazole(2.3 g, 6.9 mmol), sodium iodide (1.55 g, 10.3 mmol), 2-aminoethanethiolhydrochloride (0.94 g, 8.3 mmol), t-BOC anhydride (2.25 g, 10.3 mmol),sodium bicarbonate (1.74 g, 20.6 mmol) in dioxane (50 mL) and water (50mL) was stirred at room temperature for 6 h. The reaction mixture wasdiluted with ethyl acetate, washed with brine, and concentrated. Theresidue was chromatographed on silica gel (1% methanol/dichloromethane),affording 2.9 g of the title compound. 1H NMR (CDCl3) δ1.42 (9H), 2.62(2H), 3.05 (2H), 3.30 (4H), 3.87 (s, 2H), 5.03 (br s, 1H), 7.50-7.85(5H), 8.80 (s, 1H).

Example 149(7R)-7-amino-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester, p-toluenesulfonic acid salt

To a solution of4-[(2-tert-butoxycarbonylaminoethyl)thiomethyl]-5-(2-phenylsulfonylethyl)thio-1,3-thiazole(260 mg, 0.55 mmol) in N,N-dimethylformamide (5 mL) was added potassiumtert-butoxide in tetrahydrofuran (1 M, 0.5 mL), and the mixture wasstirred at room temperature for 30 min.(7R)-7-amino-3-methanesufonyloxy-3-cephem-4-carboxylate, diphenylmethylester (freed from its p-toluenesulfonic acid salt form just prior touse, 314 mg, 0.5 mmol) was added and stirred for 1 h. The mixture wasquenched with aqueous sodium bicarbonate, extracted with ethyl acetate,and then concentrated. The crude was chromatographed on silica gel,affording 185 mg of the free base, which was then treated withp-toluenesulfonic acid (52 mg) in ethyl acetate. Removal of the solventyielded the title compound. 1H NMR of the free base (CDCl3) δ1.42 (9H),2.64 (2H), 3.35 (4H), 3.94 (2H), 4.76 (1H), 4.92 (1H), 5.02 (br s, 1H),6.98 (1H), 6.20-7.60 (10H), 8.93 (s, 1H).

Example 150(7R)-7-[(Z)-2-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester

To a solution of(7R)-7-amino-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester, p-toluenesulfonic acid salt (232 mg, 0.28 mmol) and2-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)aceticacid (173 mg, 0.33 mmol) in anhydrous tetrahydrofuran (30 mL) weresequentially added phosphorous oxychloride (0.04 mL, 0.41 mmol) anddiisopropylethylamine (0.17 mL, 0.97 mmol) at -30° C. The resultingsolution was stirred for 1.5 h and the reaction was quenched with 0.1%hydrochloric acid. The mixture was extracted with ethyl acetate andconcentrated. The residue was then immediately chromatographed on silicagel (0.5% methanol/dichloromethane), affording 170 mg of the titlecompound. 1H NMR (CDCl3) δ1.37 (9H), 1.45 (9H), 2.63 (2H), 3.27 (4H),3.85 (q, 2H), 5.12 (d, 1H, J=5), 5.21 (br s, 1H), 6.09 (1H), 6.97 (s,1H), 7.12 (d, 1H, J=8), 7.22-7.42 (25H), 7.53 (1H), 7.80 (d, 1H, J=8),8.89 (s, 1H).

Example 151(7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

To a mixture of(7R)-7-[(Z)-2-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester (91 mg, 0.08 mmol) and phenol (200 mg) at 45° C. wasadded methanesulfonic acid (17 mg, 0.18 mmol), and the mixture wasstirred for 1.5 h. After cooling, diisopropyl ether was added. Theresulting precipitate was collected, dissolved in water, and filtered.The filtrate was subjected to reverse phase chromatography on Amberchromcolumn (from 0 to 50% water (containing 0.1% trifluoroaceticacid)/acetonitrile) to give 18 mg of the title compound. 1H NMR (D₂ O)δ2.91 (2H), 3.30 (2H), 3.46 (d, 1H, J=18), 3.60 (d, 1H, J=18), 4.12(2H), 5.35 (1H), 5.88 (1H), 7.02 (d, 1H, J=8), 7.17 (d, 1H, J=8), 7.98(t, 1H, J=8), 9.20 (s, 1H).

Example 152 4-Hydroxymethyl-3-mercaptopyridine

To a suspension of 3-mercaptoisonicotinic acid (1.80 g, 11.6 mmol) indry THF (70 mL) was added slowly borane in THF (52 ml, 1M, 52 mmol) andthe reaction mixture was stirred for 30 min. The solvent was evaporatedunder reduced pressure, methanol (40 mL) was added, and after gasevolution stopped, concentrated hydrochloric acid (3.6 mL) was added.The solution was filtered and the filtrate was evaporated to dryness.The residue was redissolved in a small volume of water, concentratedaqueous ammonia (3.6 ml) was added and the reaction mixture wasevaporated to dryness. After overnight drying under vacuum, aquantitative yield of the title product was obtained, which was used forthe next step without purification. 1H NMR (CD₃ OD) δ4.786 (s, 2H), 8.13(d, 2H, J=6), 8.60 (d, 2H, J=6), 8.70 (s, 1H).

Example 153 3-(Triphenylmethylthio)-4-hydroxymethylpyridine

4-hydroxymethyl-3-mercaptopyridine (100 mg, 0.71 mmol) was dissolved inDMF (5 mL) and diisipropylethylamine (0.12 mL, 0.71 mmol) was addedfollowed by triphenylmethylchloride (197 mg, 0.71 mmol). After 30 min.the reaction mixture was partitioned between water and ethyl acetate,and the organic layer was thoroughly washed with water and dried withanhydrous sodium sulfate. Purification by radial chromatography onsilica gel produced pure title material (67 mg, 25% yield).

1H NMR (CDCl₃) δ4.17 (s, 2H), 7.20-7.40 (m, 15H), 7.42 (d, 1H, J=6),8.22 (s, 1H), 8.40 (d, 1H, J=6).

Example 1544-{[(2-N-tert-butoxycarbonylaminoethyl)thio]methyl}-3-(triphenylmethylthio)pyridine

A solution of thionyl chloride (126 mg, 1.05 mmol) in dry DMF (2 ml) wasstirred for 30 min. at room temperature. This solution was thencannulated into a solution of3-(triphenylmethylthio)-4-hydroxymethylpyridine (270 mg, 0.70 mmol) inDMF (2 ml) at room temperature and the reaction was stirred for 30 min.2-(N-t-butoxycarbonylaminoethyl)thiol (187 mg, 1.05 mmol) and powderedpotassium carbonate (486 mg, 3.52 mmol) were added to the reactionmixture and stirring was continued for an additional 30 min. Thereaction was partitioned between water and ethyl acetate and the organiclayer was thoroughly washed with water and dried. After removing thesolvent under reduced pressure the residue was purified by radialchromatography on silica gel (hexane/ethyl acetate--4/1) to yield thetitle material as an off-white solid (220 mg, 58%).

1H NMR (CDCl₃) δ1.46 (s, 9H), 2.42 (t, 2H, J=6), 3.18 (s, 2H), 3.23 (m,2H), 4.80 (br.s, 1H), 7.10 (d, 1H, J=6), 7.20-7.45 (m, 15H), 8.28 (m,2H).

Example 155 4-[(2-N-tert-butoxycarbonylaminoethylthio)methyl]-3-(tert-butoxycarbonylthio)pyridine

To4-(2-N-t-butoxycarbonylaminoethylthiomethyl)-3-(triphenylmethylthio)pyridine(790 mg, 1.45 mmol) and triethylsilane (2 mL, 12.5 mmol) dissolved inmethylene chloride (15 mL) was added trifluoroacetic acid (15 mL). After1 hour stirring at room temperature, the reaction mixture was evaporatedto dryness under reduced pressure and the residue was dissolved in drytetrahydrofuran (10 mL). To this solution diisopropylethylamine (1.40mL, 7.84 mmol) was added, followed by di-t-butyldicarbonate (1280 mg,5.88 mmol), and after 3 hr. reaction at room temperature the solvent wasremoved at reduced pressure. Purification on silica gel by radialchromatography (methylene chloride/methanol--50/1) produced pure titleproduct as a yellow foam (460 mg, 79%).

1H NMR (CDCl₃) δ1.42 (s, 9H), 1.50 (s, 9H), 2.58 (t, 2H, J=6), 3.23 (t,2H, J=6), 3.84 (s, 2H), 4.95 (br.s, 1H), 7.42 (d, 1H, J=6), 8.58 (d, 1H,J=6), 8.70 (s, 1H).

Example 1564-(2-N-t-butoxycarbonylaminoethylthiomethyl)-3-mercaptopyridine

To a solution of4-(2-N-t-butoxycarbonylaminoethylthiomethyl)-3-(tert-butoxycarbonylthio)pyridine(415 mg, 1.04 mmol) in methanol (2 ml) was added under nitrogen amethanolic solution of sodium methoxide (1.04 mL, 1.0 M), and thereaction mixture was heated at 50° for 30 min. After evaporating thesolvent in vacuum the reaction was partitioned between water and ethylacetate with addition of acetic acid (125 mg, 2.08 mmol). Evaporation ofthe solvent under reduced pressure yielded the title material (298 mg,96% yield) which was used in the next step without further purification.1H NMR (CD₃ OD) δ1.42 (s, 9H), 2.60 (m, 2H), 3.20 (m, 2H), 4.0 (s, 2H),4.95 (br.s, 1H), 7.52 (d, 1H, J=6), 8.00 (d, 1H, J=6), 8.40 (s, 1H).

Example 157(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-[4-(N-tert-butoxycarbonylaminoethylthio)pyrid-3-ylthio]-3-cephem-4-carboxylate,diphenylmethyl ester

To a solution of4-(2-N-t-butoxycarbonylaminoethylthiomethyl)-3-mercaptopyridine (298 mg,0.99 mmol) in ethyl acetate (5 ml) was added(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-methanesulfonyloxy-3-cephem-4-carboxy latediphenylmethyl ester. After 1 hour stirring at room temperature thereaction was partitioned between ethyl acetate and dilute sodiumbicarbonate solution (30 mL, 1%). The organic layer was dried overanhydrous sodium sulfate and was evaporated under reduced pressure. Theresidue was purified by radial chromatography on silica gel to yieldpure title product (366 mg, 33%).

1H NMR (CD₃ OD) δ1.40 (s, 9H), 2.50 (m, 2H), 3.20 (m, 2H), 3.12 (d, 1H,J=16) 3.80 (s, 2H), 3.38 (d, 1H, J=16), 5.25 (d, 1H, J=6), 6.00 (d, 1H,J=6), 7.00 (s, 1H),7.20-7.40 (m, 25H), 7.42 (d, 1H, J=6) 8.40 (d, 1H,J=6), 8.50 (s, 1H).

Example 158(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-aminoethylthiopyrid-3-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

To a suspension of(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(triphenylmethoxyimino)acetamido]-3-[4-(N-tert-butoxycarbonylaminoethylthio)pyrid-3-ylthio]-3-cephem-4-carboxylate,diphenylmethyl ester (342 mg, 0.31 mmol) in methylene chloride (3.5 mL)was added triethylsilane (1.7 ml, 4.06 mmol) followed by addition oftrifluoroacetic acid (4.5 mL). After stirring at room temperature for 1hr. the reaction was cooled to 0° and isopropyl ether (30 mL) was added.Stirring was continued at 0° for 10 min and the resulting precipotatewas filtered and washed thoroughly with diisopropyl ether and dried invacuum, yielding the title deprotected cephem bis-trifluoroacetate salt(222 mg, 85%).

1H NMR (D₂ O) δ2.90 (t, 2H, J=6), 3.31 (t, 2H, J=6), 3.43 (d, 1H, J=17),3.85 (d, 1H, J=17), 4.18 (s, 2H), 5.43 (d, 1H, J=4.5), 5.97 (d, 1H,J=4.5), 8.10 (d, 1H, J=6), 8.66-8.68 (m, 2H).

Using substantially the same methods as described in the examples above,the following additional examples were prepared:

Example 159(7R)-7-[(Z)-2-(2-amino-5-chloropyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 160(7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(2-(2-aminoethylthiomethyl)phenylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 161(7R)-7-[(Z)-2-(2-amino-5-chloropyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(2-(2-aminoethylthiomethyl)phenylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 162(7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-iso-thiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 163(7R)-7-[(Z)-2-(2-amino-5-chloropyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-iso-thiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 164(7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-aminoethylthiomethyl)-1,2,5-thiadiazol-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 165(7R)-7-[(Z)-2-(2-amino-5-chloropyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(3-(2-aminoethylthiomethyl)-1,2,5-thiadiazol-4-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt Example 166(7R)-7-[(Z)-2-(2-aminopyrid-6-yl)-2-(hydroxyimino)acetamido]-3-(4-(2-aminoethylthiomethyl)-2-chloro-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,trifluoroacetic acid salt

To a mixture of(7R)-7-[(Z)-2-(2-tert-butoxycarbonylaminopyrid-6-yl)-2-(triphenylmethoxyimino)acetamido]-3-(4-(2-tert-butoxycarbonylaminoethylthiomethyl)-1,3-thiazol-5-ylthio)-3-cephem-4-carboxylate,benzhydryl ester (91 mg, 0.08 mmol) and phenol (200 mg) at 45° C. wasadded methanesulfonic acid (17 mg, 0.18 mmol), and the mixture wasstirred for 1.5 h. After cooling, diisopropyl ether was added. Theresulting precipitate was collected, dissolved in water, and filtered.The filtrate was subjected to reverse phase chromatography on Amberchromcolumn (from 0 to 50% water (containing 0.1% trifluoroaceticacid)/acetonitrile) to give 18 mg of the title compound. 1H NMR (D₂ O)δ2.91 (2H), 3.30 (2H), 3.46 (d, 1H, J=18), 3.60 (d, 1H, J=18), 4.12(2H), 5.35 (1H), 5.88 (1H), 7.02 (d, 1H, J=8), 7.17 (d, 1H, J=8), 7.98(t, 1H, J=8), 9.20 (s, 1H).

Thus, it will be appreciated that the compounds, methods andcompositions of the invention are effective against various β-lactamresistant strains of bacteria which pose an increasing health risk tosociety.

One skilled in the art would readily appreciate that the presentinvention is well adapted to carry out the objects and obtain the endsand advantages mentioned, as well as those inherent therein. Themolecular complexes and the methods, procedures, treatments, molecules,specific compounds described herein are presently representative ofpreferred embodiments are exemplary and are not intended as limitationson the scope of the invention. Changes therein and other uses will occurto those skilled in the art which are encompassed within the spirit ofthe invention are defined by the scope of the claims.

It will be readily apparent to one skilled in the art that varyingsubstitutions and modifications may be made to the invention disclosedherein without departing from the scope and spirit of the invention.

All patents and publications mentioned in the specification areindicative of the levels of those skilled in the art to which theinvention pertains. All patents and publications are herein incorporatedby reference to the same extent as if each individual publication wasspecifically and individually indicated to be incorporated by reference.

The invention illustratively described herein suitably may be practicedin the absence of any element or elements, limitation or limitationswhich is not specifically disclosed herein. Thus, for example, in eachinstance herein any of the terms "comprising", "consisting essentiallyof" and "consisting of" may be replaced with either of the other twoterms. The terms and expressions which have been employed are used asterms of description and not of limitation, and there is no intentionthat in the use of such terms and expressions of excluding anyequivalents of the features shown and described or portions thereof, butit is recognized that various modifications are possible within thescope of the invention claimed. Thus, it should be understood thatalthough the present invention has been specifically disclosed bypreferred embodiments and optional features, modification and variationof the concepts herein disclosed may be resorted to by those skilled inthe art, and that such modifications and variations are considered to bewithin the scope of this invention as defined by the appended claims.

In addition, where features or aspects of the invention are described interms of Markush groups, those skilled in the art will recognize thatthe invention is also thereby described in terms of any individualmember or subgroup of members of the Markush group. For example, if X isdescribed as selected from the group consisting of bromine, chlorine,and iodine, claims for X being bromine and claims for X being bromineand chlorine are fully described.

Other embodiments are within the following claims.

What is claimed:
 1. A compound of the formula: ##STR15## or apharmaceutically acceptable salt thereof wherein R¹ is selected from thegroup consisting of --NHC(O)ZR³, --NR⁴ R⁵, and ##STR16## Z is selectedfrom the group consisting of --CH₂ (X)_(m) --, --C(NOR⁶)--, --CH(OR⁷)--,--C(CHCO₂ R⁸)-- and --CH(NR⁹ R¹⁰)--;X is selected from the groupconsisting of oxygen and sulfur; m is selected from the group consistingof 0 and 1; R³ is selected from the group consisting of cyano,optionally substituted alkyl, optionally substituted aryl, optionallysubstituted heterocycle, optionally substituted heteroaralkyl and(CH₂)_(n) T,wherein said alkyl is optionally substituted withsubstituents selected from the group consisting of hydroxyl, bromo,fluoro, chloro, iodo, mercapto, cyano, alkylthio, carboxyl,alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl, carboxamido,isothioureido, amidino, and guanidino; wherein said aryl is substitutedwith substituents selected from the group consisting of hydroxyl, bromo,fluoro, chloro, iodo, mercapto, cyano, alkylthio, carboxyl,alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl, and carboxamido; whereinsaid heterocycle is selected from the group consisting of furyl,thienyl, imidazolyl, indolyl, pyridinyl, thiadiazolyl, thiazolyl,piperazinyl, dibenzfuranyl, dibenzthienyl, pyrimidinyl, and pyridazinyl;and wherein said heterocycle and said heteroaryl portion of saidheteroaralkyl is each independently and optionally substituted withsubstituents selected from the group consisting of hydroxyl, bromo,fluoro, chloro, iodo, mercapto, cyano, alkylthio, carboxyl, oxo,alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl, and carboxamido; n is 1to 6, T is selected from the group consisting of amino, amidino (C- orN-linked), guanidino, and isothioureido, optionally substituted byalkyl, aryl, hydroxyl, or amino; R⁴ -R⁷ are independently selected fromthe group consisting of hydrogen, optionally substituted alkyl,optionally substituted aryl and, acyl;wherein said alkyl is optionallysubstituted with substituents selected from the group consisting ofhydroxyl, bromo, fluoro, chloro, iodo, mercapto, cyano, alkylthio,carboxyl, alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl, carboxamido,isothioureido, amidino, and guanidino; and wherein said aryl issubstituted with substituents selected from the group consisting ofhydroxyl, bromo, fluoro, chloro, iodo, mercapto, cyano, alkylthio,carboxyl, alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl, andcarboxamido; R⁸ is selected from the group of hydrogen, optionallysubstituted alkyl, optionally substituted aryl, and acyl;wherein saidalkyl is optionally substituted with substituents selected from thegroup consisting of hydroxyl, bromo, fluoro, chloro, iodo, mercapto,cyano, alkylthio, carboxyl, alkoxycarbonyl, alkenyl, nitro, amino,alkoxyl, carboxamido, isothioureido, amidino, and guanidino; and whereinsaid aryl is substituted with substituents selected from the groupconsisting of hydroxyl, bromo, fluoro, chloro, iodo, mercapto, cyano,alkylthio, carboxyl, alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl, andcarboxamido; R⁹ and R¹⁰ are selected independently from the groupconsisting of hydrogen, optionally substituted alkyl, acyl, andoptionally substituted heterocyclecarbonyl;wherein said alkyl isoptionally substituted with substituents selected from the groupconsisting of hydroxyl, bromo, fluoro, chloro, iodo, mercapto, cyano,alkylthio, carboxyl, alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl,carboxamido, isothioureido, amidino, and guanidino, and wherein theheterocycle portion of said heterocyclecarbonyl is optionallysubstituted with substituents selected from the group consisting ofhydroxyl, bromo, fluoro, chloro, iodo, mercapto, cyano, alkylthio,carboxyl, oxo, alkoxycarbonyl, alkenyl, nitro, amino, alkoxyl, andcarboxamido; R² is selected from the group consisting of hydrogen,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted aryl, optionally substituted heterocycle, optionallysubstituted aralkyl, optionally substituted heteroaralkyl, andtrialkylsilyl; or when R¹² is a cationic group R² is not present and theCO₂ group to which it would be attached bears a negative charge;whereinsaid alkyl and said alkenyl are each independently and optionallysubstituted with substituents selected from the group consisting ofhydroxyl, bromo, fluoro, chloro, iodo, mercapto, cyano, alkylthio,carboxyl, alkoxycarbonyl, nitro, amino, alkoxyl, carboxamido,isothioureido, amidino, and guanidino;, wherein said aryl and said arylportion of said aralkyl is each independently and optionally substitutedwith substituents selected from the group consisting of hydroxyl, bromo,fluoro, chloro, iodo, mercapto, cyano, alkylthio, carboxyl,alkoxycarbonyl, nitro, amino, alkoxyl, and carboxamido; wherein saidheterocycle is selected from the group consisting of furyl, thienyl,imidazolyl, indolyl, pyridinyl, thiadiazolyl, thiazolyl, piperazinyl,dibenzfuranyl, dibenzthienyl, pyrimidinyl, and pyridazinyl; and whereinsaid heterocycle and said heteroaryl portion of said heteroaralkyl iseach independently and optionally substituted with substituents selectedfrom the group consisting of hydroxyl, bromo, fluoro, chloro, iodo,mercapto, cyano, alkylthio, carboxyl, oxo, alkoxycarbonyl, alkenyl,nitro, amino, alkoxyl, and carboxamido; G, H, J, L and M are carbon, ornitrogen;wherein the specific juxtaposition of groups G, H, J, L and Mforms a heterocyclic group selected from the group consisting of##STR17## R¹¹ is selected from the group consisting of hydrogen,halogen, optionally substituted alkyl, optionally substituted alkoxy,hydroxyl, amino, cyano, optionally substituted hydroxyalkyl, optionallysubstituted carboxamidoalkyl, and optionally substituted aminoalkyl;wherein said alkyl, and said alkyl portion of said alkoxy, hydroxyalkyl,carboxamidoalkyl, and aminoalkyl is each independently and optionallysubstituted with substituents selected from the group consisting ofbromo, fluoro, chloro, iodo, mercapto, cyano, alkylthio, carboxyl,alkenyl, nitro, isothioureido, amidino, and guanidino; alk₁ and alk₂ arealkyl groups and are independently and optionally substituted with asubstitutent selected from the group consisting of alkyl, hydroxyl,optionally substituted amino, alkoxy, hydroxyalkyl, and optionallysubstituted carboxamide; p is 0, 1, or 2; R⁹⁹ is selected from the groupconsisting of sulfur, SO, SO₂, NH, N-alkyl, oxygen, C═C (cis or trans),and C.tbd.C; q is 1; r is 0, 1, 2 or 3; R¹² is NR¹³ R¹⁴, ##STR18## R¹³-R¹⁶ are independently selected from the group consisting of, hydroxy,amino, amidino, alkyl, cycloalkyl, acyl, aminoacyl, and phosphoryl; andR¹³ -R¹⁶ are independently selected from the group consisting ofhydrogen, hydroxy, amino, amidino, optionally substituted alkyl,cycloalkyl, acyl, aminoacyl, and phosphoryl;wherein said alkyl isoptionally substituted with substituents selected from the groupconsisting of bromo, fluoro, chloro, iodo, mercapto, cyano, alkylthio,heterocycle, aryl, heteroaryl, nitro, alkoxyl, carboxamido,isothioureido, amidino, and guanidino; R¹⁷ is hydrogen or optionallysubstituted alkyl;wherein said alkyl is optionally substituted withsubstituents selected from the group consisting of bromo, fluoro,chloro, iodo, mercapto, cyano, alkylthio, heterocycle, aryl, heteroaryl,nitro, alkoxyl, carboxamido, isothioureido, amidino, and guandino. 2.The compound or salt of claim 1, wherein R¹ is --NHC(O)ZR³.
 3. Thecompound or salt of claim 2, wherein R³ is optionally substituted arylor an optionally substituted heterocycle.
 4. The compound or salt ofclaim 3, wherein R³ is selected from the group consisting of optionallysubstituted phenyl, optionally substituted thienyl, optionallysubstituted furanyl, 2-aminothiazol-4-yl, 2-amino-5-chlorothiazol-4-yl,2-amino-pyrid-6-yl, 2-amino-5-chloro-pyrid-6-yl, and2-aminothiadiazol-4-yl.
 5. The compound or salt of claim 4, wherein Z is--C(NOR⁶)--, and R⁶ is selected from the group consisting of hydrogenand optionally substituted alkyl.
 6. The compound or salt of claim 5,wherein R⁶ is and R³ is selected from the group consisting of phenyl,2-thienyl, 2-furyl; 2-aminothiazol-4-yl, 2-amino-5-chlorothiazol-4-yl,2-amino-pyrid-6-yl, 2-amino-5-chloro-pyrid-6-yl, and2-aminothiadiazol-4-yl.
 7. The compound or salt of claim 6, wherein R³is 2-aminothiazol-4-yl, 2-amino-pyrid-6-yl, 2-amino-5-chloro-pyrid-6-yl,or 2-amino-5-chlorothiazol-4-yl.
 8. The compound or salt of claim 7,wherein said compound is active against methicillin-resistantStaphylococcal or ampicillin-resistant Enterococcal bacteria, asdemonstrated by a lower minimum inhibitory concentration thanmethicillin or ampicillin respectively, wherein said bacteria areselected from the group consisting of S. aureus Col (Meth^(R))(lac-), S.aureus 76 (Meth^(R))(lac+), S. aureus ATCC 33593(Meth^(R)), S. aureusSpain #356 (Meth^(R)), S. haemolyticus 05 (Meth^(R)), E. faecalis ATCC29212, E. faecium ATCC 35667, E. faecium VanA (Van^(R)), E. faecalisVanB (Van^(R)), and E. faecium A491 (Amp^(R)).
 9. A method of treating amammal suffering from a methicillin-resistant Staphylococcal, orampicillin-resistant Enterococcal bacterial infection, comprisingadministering to such mammal a therapeutically effective amount of acompound or salt of any one of claims 1-8.
 10. The method of claim 9,wherein said mammal is infected with a methicillin-resistantStaphylococcal or ampicillin-resistant Enterococcal organism.
 11. Anantibacterial composition for treating a methicillin-resistantStaphylococcal or ampicillin-resistant Enterococcal bacterial infection,comprising a therapeutically effective amount of a compound or salt ofany one of claims 1-8 in a pharmaceutically acceptable carrier.
 12. Thecomposition of claim 11, wherein said bacteria is amethicillin-resistant Staphylococcal or ampicillin-resistantEnterococcal organism.
 13. The compound or salt of claim 1, wherein:R¹is --NHC(O)ZR³ ;Z is --C(NOR⁶)R⁶ is hydrogen; R³ is an optionallysubstituted heterocycle; R² is hydrogen or when R¹² is a cationic groupR² is not present and the CO₂ group to which it would be attached bearsa negative charge; G,H, L and M are carbon and J is nitrogen or G, L, M,and J are carbon and H is nitrogen; alk₁ is CH₂ ; p is 0 or 1; R⁹⁹ issulfur; q is 1; alk₂ is CH₂ r is 2 or 3; R¹² is NH₂ ; and there are zeroR¹¹ groups and one group.
 14. The compound or salt of claim 13, whereinR³ is selected from the group consisting of 2-aminothiazol-4-yl,2-amino-5-chlorothiazol-4-yl, and 2-aminothiadiazol-4-yl.
 15. Thecompound or salt of claim 1, wherein said compound or salt is(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 16. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 17. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2-aminoethylthio)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 18. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2R-2-amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylic acid.
 19. The compound orsalt of claim 1, wherein said compound or salt is(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2S-2-amino-2-carboxamidoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 20. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2R-2-amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 21. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-[3-(2S-2-amino-3-hydroxypropylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 22. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 23. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-fluoroethoxyimino)acetamido]-3-[3-(2-aminoethylthiomethyl)pyrid-4-ylthio]-3-cephem-4-carboxylicacid.
 24. The compound or salt of claim 1, wherein said compound or saltis(7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-(4-aminoethylthiopyrid-3-ylthio)-3-cephem-4-carboxylicacid.
 25. A method of treating a mammal suffering from amethicillin-resistant Staphylococcal or ampicillin-resistantEnterococcal bacterial infection, comprising administering to saidmammal a therapeutically effective amount of a compound or salt of anyone of claims 13-24.
 26. The method of claim 25, wherein said mammal isinfected with a methicillin-resistant Staphylococcal orampicillin-resistant Enterococcal organism.
 27. An antibacterialcomposition for treating a methicillin-resistant Staphylococcal orampicillin-resistant Enterococcal bacterial infection comprising acompound or salt of any one of claims 13-24 in a pharmaceuticallyacceptable carrier.
 28. A compound of claim 13 of the formula: ##STR19##or a pharmaceutically acceptable salt thereof, wherein R³⁴ is halogen orhydrogenR⁵³ is selected from the group consisting of NH₂ and protectedamino; and R², R⁶, G, H, J, L, M, R¹¹, alk₁, p, R⁹⁹, q, alk₂, r, and R¹²are as defined in claim 13 above.
 29. The compound or salt of claim 28,whereinR² is hydrogen, or when R¹² is a cationic group R² is not presentand the CO₂ group to which it would be attached bears a negative charge;R³⁴ is chlorine or hydrogen; R⁶ is hydrogen; G, H, J, L, and M areselected to form 2-pyridyl, 3-pyridyl, or 4-pyridyl; R¹¹ is hydrogen,halogen or alkyl; there is one group; alk₁ is CH₂ ; p is 0 or 1; R⁹⁹ issulfur; q is 1; alk₂ is CH₂ optionally substituted with hydroxy,hydroxyalkyl or optionally substituted carboxamide; r is 2; and R¹² isNH₂.
 30. The compound or salt of claim 29, wherein R³⁴ is chlorine, R⁶is hydrogen, and G, H, J, L, and M are selected to form 4-pyridyl. 31.The compound or salt of claim 29, wherein R³⁴ is chlorine, R⁶ ishydrogen, and G, H, J, L, and M are selected to form 3-pyridyl.